Insilico Analysis of Sesamum indicum Seed Proteins

 

Rangarajan Narasimhan* and Ambilly Mohan

Dept. of Biochemistry, Sri Sankara Arts and Science College, Enathur, Kanchipuram-631561, Tamil Nadu, India

*Corresponding Author E-mail: n.rangarajan@yahoo.co.in

 

ABSTRACT:

Many infectious diseases have been known to be treated with herbal remedies throughout the history of mankind. The use of herbal medicines in science in an evidence based manner is called phytotherapy. Natural products, either as pure compounds or standardized plant extracts provide unlimited opportunities for new drug release because of chemical diversity. In the present study, an attempt has been made to quantitate proteins in sesame seeds, to separate proteins by SDS PAGE and to analyse the molecular weight by marker protein. The current study also aimed to conduct multiple sequence alignment, similarity searches and secondary structure prediction of the major globulins in sesame seeds (7S globulin and 11S globulin) using bioinformatics tools. The study of protein shows that the sequences of 7Sglobulin and 11S globulin of Sesamum indicum have greater sequence homology with 7S globulin of Elaeis guineensis var tenera and 11S storage globulin of Coffea arabica respectively. The results also reveal the presence of alpha helix, extended strand and random coils in7S and 11S globulin with a high occurrence of random coils in both the globulin subunits.

 

KEY WORDS: Sesamum indicum, globulin, BLAST, Multiple sequence alignment, FASTA.

 

 

INTRODUCTION:

Over the past decade, herbal medicines have been accepted universally, and they have an impact on both world health and international trade. Hence, medicinal plants continue to play an important role in the healthcare system of a large number of the world’s population1. Traditional medicine is widely used in India. Even in USA, use of plants and phytomedicines has increased dramatically in the last two decades. The herbal products have been classified under ‘dietary supplements’ and are included with vitamins, minerals, amino acids and other products intended to supplement the diet2.

 

Medicinal plants and herbal preparations have recently received considerable attention and have been found to be promising choice over modern synthetic medicines, in a number of studies. In developing countries, all over the world, 80% of population continues to use traditional medicine in primary medical problems3. Research carried out in last few decades has validated several such claims of use of traditional medicinal plants. Pumpkin is one such plant that is frequently being used as food as well as traditional medicine for long days4.

 

Sesame (Sesamum indicum) is one of about 15 species of herbaceous plants of the genus ‘Sesamum’ native to Africa and Asia and is the most widely cultivated species for its nutritious seeds and oil. Historians believe that the original homeland of the sesame seed is the Indian subcontinent. The seed has been called the “Queen of oilseed crops” because of high yield of oil and the quality of seed, oil and meal. Sesame oil is non-drying oil, highly stable, rarely turning rancid in hot climates5.

Scientific Classification

Kingdom

:

Plantae

Division

:

Angiosperms

Order

:

Lamiales

Family

:

Pedaliaceae

Genus 

:

Sesamum

Species

:

S. indicum.

 

Binomial name:

Sesamum indicum L

 

Sesame is a herbaceous annual and tropical plant which reaches height of 1-2 meters. The leaves have varied shapes, they are either oval or lanceolate and fluffy on both sides. The flowers are either violet or white and at their end, one can find pods of 3cm containing many seeds. These are small and have a length of 3.5 mm. Their colour varies from yellowish-white to red, brown or black6. Lignans and lignan glycosides present in sesame appear to be the important functional components. The main sesame lignans are sesamin and sesamolin which are found in sesame oil7.

 

MATERIALS AND METHODS:

Estimation of Proteins by Lowry’s method.

The solid part left behind after soxhelet extraction was dissolved in distilled water at a concentration of 1g/100ml. the protein level was determined by Lowry’s method8.The values were expressed as mg% of protein.

 

Chemicals Used:

Acrylamide and bisacrylamide, TEMED, Coomassie brilliant blue, bromophenol blue, ammonium persulfate, sodium dodecyl sulphate. Dithiothreitol, Tris, Glycine were obtained from standard chemical companies like Qualigens, SRL etc.

 

SDS PAGE Analysis of Proteins:

The solid residue left behind in the thimble after extraction in soxhelet apparatus was used for analysis of proteins in SDS PAGE in the concentration of 0.03mg/ml.  Molecular weight Marker Protein of 6.5-70kDa was run along with the samples.

 

Protein sequence analysis using bioinformatic tools:

Retrieval of protein sequence:

The protein sequence of 7S and 11S globulin of Sesamum indicum were retrieved from UniprotKB Database. Their accession numbers were Q9AUD2_SESIN and Q9AUDO – SESIN respectively.  The website utilized was http://www.uniprot.org.

 

Similarity studies:

Similarity Studies were carried out using different alignment methods.

 

Pair wise Alignment:

It refers to the comparison of two sequences.

 

 

Local Sequence Alignment:

BLAST (Basic Local Alignment Search Tool) in NCBI is utilized to find regions of sequence similarity, which will yield functional and evolutionary clues about structure and function of query sequence9. Protein sequences of 7s globulin and 11s globulin of Sesamum indicum obtained in FASTA format were submitted individually in BLASTP with low complexity region filter for 11s globulin and default parameters for 7s globulin. Individual sequences were compared with sequences in non redundant protein (nr) database using protein BLAST and a graphic overview of alignments were obtained.

 

 

Multiple Sequence Alignment and Phylogenetic Analysis:

Multiple sequence alignment is an alignment of three or more sequences with gap utilized for conserved region identification and phylogenetic analysis.  ClustalW was used to find multiple sequence alignment of 7s and 11s globulin of Sesamun indicum with globulins of other plant species. The protein sequences for 7s globulin and 11s globulin of Sesamum indicum retrieved from uniprot database were converted to FASTA format. Similarly globulin sequences of other plant species in Uniprot database were retrieved in FASTA format. Multiple sequence analysis was carried by comparing the globulin sequences of Sesamum indicum separately with other globulin sequences in Uniprot using clustalW with default parameters.

 

STRUCTURAL ANALYSIS:

Primary structure analysis

Molecular weight, isoelectric point and amino acid  composition of 7S and 11S globulin of Sesamum indicum was identified using Protparam tool of Expasy server10 .

 

Secondary structure Prediction

Secondary structures of 7s and 11s globulin sequences of Sesamum indicum was predicted using GOR method server11.

 

RESULTS AND DISCUSSION:

                     

Figure 1. Separation of protein subunits of sesamum indicum seed extracts by SDS- PAGE

Lane 1 - Protein molecular weight marker.

Lane 2 - Methanolic extract of sesamum indicum seed.

Lane 3 - Hexane extract of sesamum indicum seed.

 

SDS PAGE ANALYSIS OF PROTEINS:

Figure 1 shows the separation of proteins present in Sesamum indicum by SDS - PAGE. Lane 1 shows the separation of molecular weight marker protein of molecular weight ranging from 6.5 - 70 kDa.  Lanes 2 and 3 gives the separation of methanolic and hexane extracts respectively. It is clear from the figure that proteins containing different subunits ranging 6.5 - 70 kDa are present in Sesamum indicum. This result correlates will with the results obtained by V. Prakash and P.K. Nandi12 by Polyacrylamide Gel Electorphoresis of total protein of Sesamum indicum. They reported that gel elctrophoresis of total protein shows a major band along with several slow and fast moving components.  The oligomeric nature of the protein is shown by the presence of 12 sub units in it.  The molecular weights of subunits of alpha globulin ranged from 8kDa - 85 kDa13.  The major globulins of Sesamum indicum are 7s and 11s globulins.  Most of the subunits of these globulins ranges from 6.5 – 75kDa.  Our results correlate with the results of Orruno E and M.R.A. Morgan. They isolated 7s globulin from sesame seeds and reported that the isolated protein was composed of atleast 8 polypeptide chains ranging from 12.4 - 65.5kDa judged by SDS PAGE analysis14.

 

 

PROTEIN SEQUENCE ANALYSIS:

SEQUENCE RETRIEVAL:

7S GLOBULIN

>tr|Q9AUD0|Q9AUD0_SESIN 7S globulin OS=Sesamum indicum PE=2 SV=1

MSCGGRLCLVLFALLLASAVVASESKDPELKQCKHQCKAQQQISKEQKEACIQACKEYIR

QKHQGEHGRGGGDILEEEVWNRKSPIERLRECSRGCEQQHGEQREECLRRCQEEYQREKG

RQDDDNPTDPEKQYQQCRLQCRRQGEGGGFSREHCERRREEKYREQQGREGGRGEMYEGR

EREEEQEEQGRGRIPYVFEDQHFITGFRTQHGRMRVLQKFTDRSELLRGIENYRVAILEA

EPQTFIVPNHWDAESVVFVAKGRGTISLVRQDRRESLNIKQGDILKINAGTTAYLINRDN

NERLVLAKLLQPVSTPGEFELFFGAGGENPESFFKSFSDEILEAAFNTRRDRLQRIFGQQ

RQGVIVKASEEQVRAMSRHEEGGIWPFGGESKGTINIYQQRPTHSNQYGQLHEVDASQYR

QLRDLDLTVSLANITQGAMTAPHYNSKATKIALVVDGEGYFEMACPHMSRSRGSYQGETR

GRPSYQRVASRLTRGTVVIIPAGHPFVAVASSNQNLQVLCFEVNANNNEKFPLAGRRNVM

NQLEREAKELAFGMPAREVEEVSRSQQEEFFFKGPRQQQQGRADA

 

11S GLOBULIN

>tr|Q9AUD2|Q9AUD2_SESIN 11S globulin OS=Sesamum indicum PE=2 SV=1

MALTSLLSFFIVVTLLIRGLSAQLAGEQDFYWQDLQSQQQHKLQARTDCRVERLTAQEPT

IRFESEAGLTEFWDRNNQQFECAGVAAVRNVIQPRGLLLPHYNNAPQLLYVVRGRGIQGT

VIPGCAETFERDTQPRQDRRRRFMDRHQKVRQFRQGDILALPAGLTLWFYNNGGEPLITV

ALLDTGNAANQLDQTFRHFFLAGNPQGGRQSYFGRPQTEKQQGETKNIFNGFDDEILADA

FGVDVQTARRLKGQDDLRGRIVRAERLDIVLPGEEEEERWERDPYSGANGLEETLCTAKL

RENLDEPARADVYNPHGGRISSLNSLTLPVLSWLRLSAEKGVLYRNGLVAPHWNLNAHSI

IYITRGSGRFQVVGHTGRSVFDGVVREGQLIIVPQNYVVAKRASQDEGLEWISFKTNDNA

MTSQLAGRLSAIRAMPEEVVMTAYQVSRDEARRLKYNREESRVFSSTSRYSWPRSSRPMS

YMPKPFEYVLDVIKSMM

SEQUENCE SIMILARITY ANALYSIS USING BLAST

7S GLOBULIN

 

 

Sequences producing significant alignments:

Accession

Description

Max score

Total score

Query coverage

E value

AAK15089.1

7S globulin [Sesamum indicum]

1201

1201

100%

0.0

AAL86739.1

48-kDa glycoprotein precursor [Corylus avellana]

514

554

84%

1e-143

ABO36677.1

vicilin [Pistacia vera]

506

506

95%

4e-141

XP_002279445.1

PREDICTED: hypothetical protein isoform 2 [Vitis vinifera]

469

469

96%

4e-130

CBI28721.1

unnamed protein product [Vitis vinifera]

466

466

68%

4e-129

AAM73729.1

vicilin-like protein [Anacardium occidentale]

454

533

97%

2e-125

AAM73730.2

vicilin-like protein [Anacardium occidentale]

454

533

97%

2e-125

CAP06319.1

convicilin [Lathyrus tingitanus]

244

244

76%

3e-62

BAD98463.1

beta-conglycinin beta subunit [Glycine max] >dbj|BAE02728.1| beta-conglycinin beta subunit [Glycine max]

243

243

65%

4e-62

P10562.1

RecName: Full=Canavalin; Flags: Precursor >emb|CAA29910.1| unnamed protein product [Canavalia gladiata]

243

243

69%

5e-62

1IPJ_A

Soybean Beta- Conglycinin Beta Homotrimers

243

243

65%

6e-62

 

11S  GLOBULIN

Sequences producing significant alignments:

Accession

Description

Max score

Total score

Query coverage

E value

AAK15087.1

11S globulin [Sesamum indicum]

882

882

100%

0.0

ABB60054.1

11S globulin precursor isoform 3 [Sesamum indicum]

491

491

89%

7e-137

ABB77213.1

11S globulin-like protein [Actinidia chinensis]

468

468

84%

8e-130

XP_002329473.1

predicted protein [Populus trichocarpa]

466

466

84%

3e-129

XP_002329472.1

predicted protein [Populus trichocarpa]

466

466

84%

3e-129

CAA57846.1

legumin precur [Magnolia salicifolia]

457

457

84%

1e-126

CAA57847.1

globulin precursor [Magnolia salicifolia]

455

455

93%

6e-126

NP_001046434.1

Os02g0248800 [Oryza sativa (japonica cultivar-group)] 

360

360

92%

3e-   97

EEC72834.1

hypothetical protein OsI_06562 [Oryza sativa Indica Group]

359

359

82%

3e-   97

XP_002533465.1

legumin B precursor, putative [Ricinus communis]

359

359

71%

4e-97

EEE56664.1

hypothetical protein OsJ_06085 [Oryza sativa Japonica Group]

358

358

83%

8e-97

BAD28254.1

glutelin precursor [Oryza sativa Japonica Group]

358

358

83%

8e-97

 

MULTIPLE   SEQUENCE ALIGNMENT

7S GLOBULIN

Scores Table

SeqA  Name

Len(aa) 

SeqB Name

Len(aa)

Score

1

tr|Q9AUD0|Q9AUD0_SESIN 

585

2

sp|P13917|7SB1_SOYBN 

427

4

1

tr|Q9AUD0|Q9AUD0_SESIN 

585

3

tr|O23211|O23211_ARATH 

499

16

1

tr|Q9AUD0|Q9AUD0_SESIN 

585

4

tr|O65042|O65042_ORYSA

174

18

1

tr|Q9AUD0|Q9AUD0_SESIN 

585

5

tr|Q9AU64|Q9AU64_ELAGV 

572

42

1

tr|Q9AUD0|Q9AUD0_SESIN 

585

6

tr|Q84NG7|Q84NG7_HORVU

224

13

1

tr|Q9AUD0|Q9AUD0_SESIN

585

7

tr|Q41727|Q41727_ZAMFU

414

32

2

sp|P13917|7SB1_SOYBN   

427

3

tr|O23211|O23211_ARATH

499

8

2

sp|P13917|7SB1_SOYBN    

427

4

tr|O65042|O65042_ORYSA

174

12

2

sp|P13917|7SB1_SOYBN      

427

5

tr|Q9AU64|Q9AU64_ELAGV

572

5

2

sp|P13917|7SB1_SOYBN   

427

6

tr|Q84NG7|Q84NG7_HORVU

224

5

2

sp|P13917|7SB1_SOYBN   

427

7

tr|Q41727|Q41727_ZAMFU

414

6

3

tr|O23211|O23211_ARATH  

499

4

tr|O65042|O65042_ORYSA 

174

13

3

tr|O23211|O23211_ARATH  

499

5

tr|Q9AU64|Q9AU64_ELAGV 

572

19

3

tr|O23211|O23211_ARATH 

499

6

tr|Q84NG7|Q84NG7_HORVU 

224

8

3

tr|O23211|O23211_ARATH 

499

7

tr|Q41727|Q41727_ZAMFU

414

23

4

tr|O65042|O65042_ORYSA 

174

5

tr|Q9AU64|Q9AU64_ELAGV

572

14

4

tr|O65042|O65042_ORYSA

174

6

tr|Q84NG7|Q84NG7_HORVU

224

4

4

tr|O65042|O65042_ORYSA 

174

7

tr|Q41727|Q41727_ZAMFU 

414

16

5

tr|Q9AU64|Q9AU64_ELAGV

572

6

tr|Q84NG7|Q84NG7_HORVU

224

11

5

tr|Q9AU64|Q9AU64_ELAGV

572

7

tr|Q41727|Q41727_ZAMFU

414

32

6

tr|Q84NG7|Q84NG7_HORVU

224

7

tr|Q41727|Q41727_ZAMFU 

414

12

 

CLUSTAL 2.0.12 multiple sequence alignment

tr|Q9AUD0|Q9AUD0_SESIN      ----MSCGGRLCLVLFALLLASAVVASESKDPELKQCKHQCKAQQQISKE 46

tr|Q9AU64|Q9AU64_ELAGV      ----MTIKPR-AFVPFLLLLSILFVS----------ATLTFSATTEDPKQ 35

tr|Q41727|Q41727_ZAMFU      ---MAHLCSLPLMAVLMLLLASACFS------------------------ 23

tr|O23211|O23211_ARATH      ----MTRFAVLPLSVLLLVLLFLCTES--------------LAKSEESEE 32

tr|O65042|O65042_ORYSA      --------------------------------------------------

sp|P13917|7SB1_SOYBN        MASILHYFLALSLSCSFLFFLS---------------------------- 22

tr|Q84NG7|Q84NG7_HORVU      -------------------------------------------------

                                                                             

tr|Q9AUD0|Q9AUD0_SESIN      QKEACIQACKEYIRQKHQGEHGRGGGDILEEEVWNRKSPIERLRECSRGC 96

tr|Q9AU64|Q9AU64_ELAGV      RLERCKQECRE----SRQGER--------------------QERRCVSQC 61

tr|Q41727|Q41727_ZAMFU      --------------------------------------------------

tr|O23211|O23211_ARATH      YDVAVPSCCG---------------------------------------- 42

tr|O65042|O65042_ORYSA      --------------------------------------------------

sp|P13917|7SB1_SOYBN        --------------------------------------------------

tr|Q84NG7|Q84NG7_HORVU      --------------------------------------------------

                                                                             

tr|Q9AUD0|Q9AUD0_SESIN      EQQHG-EQREECLRRCQEEYQREKGRQDDDNPTDPEKQYQQCRLQCRRQG 145

tr|Q9AU64|Q9AU64_ELAGV      EERYERERREQEERKGQGE---ERGRRE-----EPEKRLEECRRECREQA 103

tr|Q41727|Q41727_ZAMFU      --------------------------------------------------

tr|O23211|O23211_ARATH      --------------------------------------------------

tr|O65042|O65042_ORYSA      --------------------------------------------------

sp|P13917|7SB1_SOYBN        --------------------------------------------------

tr|Q84NG7|Q84NG7_HORVU      --------------------------------------------------

                                                                             

tr|Q9AUD0|Q9AUD0_SESIN      EGGGFSREHCERRREEKYREQQGREGGRGEMYEGREREEEQEEQGRGRIP 195

tr|Q9AU64|Q9AU64_ELAGV      ERR--ERRECEKRCEEEYKEHRGRSKDKEEGEEGRG------EKRRESDP 145

tr|Q41727|Q41727_ZAMFU      -------------------------------------------ELEIEDP 30

tr|O23211|O23211_ARATH      ----------------------------------------------FSSP 46

tr|O65042|O65042_ORYSA      --------------------------------------------------

sp|P13917|7SB1_SOYBN        ----------------------------------------------DSVT 26

tr|Q84NG7|Q84NG7_HORVU      -----------------------------------------------MGK 3

                                                                             

 

tr|Q9AUD0|Q9AUD0_SESIN      YVFEDQHFITGFRTQHGRMRVLQKFTD--RSELLRGIENYRVAILEAEPQ 243

tr|Q9AU64|Q9AU64_ELAGV      YFFDEESFLHRVRTEHGHVRVLRNFLE--KSKLLLGVANYRVAILETNPN 193

tr|Q41727|Q41727_ZAMFU      YVFDQRSFVTTVQCKAGQIRALPNFSAGGRCELPRGLGDYSVAQISLEPR 80

tr|O23211|O23211_ARATH      LLIKKDQWKPIFETKFGQISTVQIGNG------CGGMGPYKIHSITLEPN 90

tr|O65042|O65042_ORYSA      ------------ETERGGEESYNILSG-------------------FDTE 19

sp|P13917|7SB1_SOYBN        PTKPINLVVLPVQNDGSTGLHWANLQKRTPLMQVPVLVDLNGNHLWVNCE 76

tr|Q84NG7|Q84NG7_HORVU      FIFFAVFLTTLVTISAAQGVLEQSLAD-----------------AQCRGE 36

                                          . .                                .

tr|Q9AUD0|Q9AUD0_SESIN      TFIVPNHWDAESVVFVAKGRGTISLVRQDRRESLNIKQGDILKINAGTTA 293

tr|Q9AU64|Q9AU64_ELAGV      TFVLPSHWDAEALLFVARGHGHITLQCQDNKATHELRRGDIMRVRAGTIV 243

tr|Q41727|Q41727_ZAMFU      SVLLPHYIEADLALYVTGGRGRVAFVHEERLVERQLRDGDVYAIAAGIPF 130

tr|O23211|O23211_ARATH      TILLPLLLHSDMVFFVDSGSGILNWVDEEAKS-TEIRLGDVYRLRPGSVF 139

tr|O65042|O65042_ORYSA      LLAESMRVSPDIARKLQGRSD---------------KRGNIVRVRRG--- 51

sp|P13917|7SB1_SOYBN        QQYSSKTYQAPFCHSTQCSRANTHQCLSCPAASRPGCHKNTCGLMSTNPI 126

tr|Q84NG7|Q84NG7_HORVU      VQAKPLLACRQILEHQLTGRAVGVRPFQAQWGARDRCCQQLESVSRGCRC 86

                                .                                  :   :     

tr|Q9AUD0|Q9AUD0_SESIN      YLINR-DNNERLVLAKLLQPVSTPGEFELFFGAGGENPESFFKSFSDEIL 342

tr|Q9AU64|Q9AU64_ELAGV      SFANRGVGNEKLVIVILLHPVATPGMFEAFVGAGGQNPESFYRSFSKRVL 293

tr|Q41727|Q41727_ZAMFU      YILNTDDSRRLFIHCLLRTQCSTTGLYESFYVVGGRNPQNVLSQFSEDVL 180

tr|O23211|O23211_ARATH      YLQSK---PDPCFGAYSSITDLMFGFDETILQSAFGVPEGIIELMRNRTK 186

tr|O65042|O65042_ORYSA      -------------------------------------GLRMLRPATERVT 64

sp|P13917|7SB1_SOYBN        TQQTG----------LGELGEDVLAIHATQGSTQQLGPLVTVPQFLFSCA 166

tr|Q84NG7|Q84NG7_HORVU      SALRG-----------------------------------MVRDYEQSMP 101

                                                                              

tr|Q9AUD0|Q9AUD0_SESIN      EAAFNTRRDRLQRIFGQ-QRQGVIVKASE--EQVRAMSRHEEGGIWPFGG 389

tr|Q9AU64|Q9AU64_ELAGV      SAAFNTREDKLERLFQK-QNKGAIIQASQ--EQIKEMSRGSEGRSWPFG- 339

tr|Q41727|Q41727_ZAMFU      QAAFNSSKAVLDPMLVSGFNRGAIIRVSR--EQMERLSRGRIKG--FGGS 226

tr|O23211|O23211_ARATH      PPLIVSETLCTPGVANTWQLQPRLLKLFAGSADLVDNKKKKEKKEKKEKV 236

tr|O65042|O65042_ORYSA      DEEMMRGANAAAAAGNGIDEAVCLMKLREN-------------------V 95

sp|P13917|7SB1_SOYBN        PSFLVQKGLPRNTQGVAGLGHAPISLPNQLASHFGLQRQFTTCLSRYPTS 216

tr|Q84NG7|Q84NG7_HORVU      PLREGRRRSSGERQQEQGCSGESTAEQQQE-------------------- 131

                                                                             

tr|Q9AUD0|Q9AUD0_SESIN      ESKGTINIYQQRPTHSNQYGQLHEVDASQYRQLRDLDLTVSLANITQGAM 439

tr|Q9AU64|Q9AU64_ELAGV      ESRRPFNLFHKRPAHSNRHGELREADSDDYPELRDLNIHVSYANISKGSM 389

tr|Q41727|Q41727_ZAMFU      EEPQPFNLLYRNPDFSNNNGEIFTADAADHRVLRRLNVGVQLLNLKPRSM 276

tr|O23211|O23211_ARATH      KKAKTFNVFESEPDFESPYGRTITINRKDLKVLKGSMVGVSMVNLTQGSM 286

tr|O65042|O65042_ORYSA      ADPMKADLYTPNG------GRITVLNSQKLPVLKLIKMSVNRGVMRRNAI 139

sp|P13917|7SB1_SOYBN        KGAIIFGDAPNNMRQFQNQDIFHDLAFTPLTITLQGEYNVRVNSIRINQH 266

tr|Q84NG7|Q84NG7_HORVU      ---------VQGGQYGSETGESQQQQGGGYHG-----VTVGRGGQQQGQM 167

                                               .                   *         

tr|Q9AUD0|Q9AUD0_SESIN      TAPHYNSKATKIALVVDGEGYFEMACPHMSRS-----RGSYQGETRGRPS 484

tr|Q9AU64|Q9AU64_ELAGV      IAPNYNTEATKISVVVGGNGDVQIVCPHISRQQEEGRRGREEEEGRGRQE 439

tr|Q41727|Q41727_ZAMFU      TAPHYDTRSTRIGIVRNGRGILELVRPQEQEQ---------QQQQQQGPT 317

tr|O23211|O23211_ARATH      MGPHWNPWACEISIVLKGAGMVRVLRSSISSN--------TSSECKN--- 325

tr|O65042|O65042_ORYSA      LAPHWNINAHAAVYATSGSAKLQVVS------------------------ 165

sp|P13917|7SB1_SOYBN        SVFPLNKISSTIVGSTSGGTMISTSTPHMVLQQSVYQAFTQVFAQQLPKQ 316

tr|Q84NG7|Q84NG7_HORVU      LCRER-PQRQQQGEGFSGEG------------------------------ 186

                                             *                               

 

tr|Q9AUD0|Q9AUD0_SESIN      ----------------YQRVASRLTRGTVVIIPAGHPFVAVASSNQNLQV 518

tr|Q9AU64|Q9AU64_ELAGV      GREEEEEEEQQQRGQHYRRVESKVSCGTTFIVPAGHPSVSVSSRNENLEV 489

tr|Q41727|Q41727_ZAMFU      ----------------YQKLRANLNPGTVFLTRPGYPSTVIASGNEALQI 351

tr|O23211|O23211_ARATH      -------------------VRFKVEEGDIFAVPRLHPMAQMSFNNDSLVF 356

tr|O65042|O65042_ORYSA      --------------------------------------------------

sp|P13917|7SB1_SOYBN        -----------------AQVKSVAPFGLCFNSNKINAYPSVDLVMDKPNG 349

tr|Q84NG7|Q84NG7_HORVU      --------------------------------------------------

                                                                             

tr|Q9AUD0|Q9AUD0_SESIN      LCFEVNANNNEKFPLAGRRNVMNQLEREAKELAFGMPAREVEEVSRSQQE 568

tr|Q9AU64|Q9AU64_ELAGV      LCFEINAKNNQRTWLAGRNNVLKQMDRVTKELAFDLPEREVDEVLNAPRE 539

tr|Q41727|Q41727_ZAMFU      LYLDNYSQGSRRQFLAGRSNVLRYLPREIKRLVFPSSAEEIEATLEAQED 401

tr|O23211|O23211_ARATH      VGFTTSAKNNEPQFLAGEDSALRMLDRQVLAASLNVSSVTIDGLLGAQKE 406

tr|O65042|O65042_ORYSA      --------NEGRPVFDG--------------------------------- 174

sp|P13917|7SB1_SOYBN        PVWRISGEDLMVQAQPGVTCLGVMNGGMQPRAEITLGARQLEENLVVFDL 399

tr|Q84NG7|Q84NG7_HORVU      --------AQQKPKVGRVRLTKVRLPTACRIEPQECSVFSTLPVLG---- 224

                                                                             

tr|Q9AUD0|Q9AUD0_SESIN      EFFFKGP-------------------RQQQQGRADA-------------- 585

tr|Q9AU64|Q9AU64_ELAGV      EVFMAGPEERG-------------RERERGEGRDGPLESILEFAGF---- 572

tr|Q41727|Q41727_ZAMFU      EVLLNAQ-----------------------QGRADQ-------------- 414

tr|O23211|O23211_ARATH      AVILECHSCAEGEIEKLKVEIERKKIDDERKRRHDERKKEEEEAKREEEE 456

tr|O65042|O65042_ORYSA      --------------------------------------------------

sp|P13917|7SB1_SOYBN        ARSRVGFSTSS--------------LHSHGVKCADLFNFANA-------- 427

tr|Q84NG7|Q84NG7_HORVU      --------------------------------------------------

tr|Q9AUD0|Q9AUD0_SESIN      -------------------------------------------

tr|Q9AU64|Q9AU64_ELAGV      -------------------------------------------

tr|Q41727|Q41727_ZAMFU      -------------------------------------------

tr|O23211|O23211_ARATH      RRKREEEEEKKRWPPQQPPQEEELRERQLPMEKEWEMEGEEES 499

tr|O65042|O65042_ORYSA      -------------------------------------------

sp|P13917|7SB1_SOYBN        -------------------------------------------

tr|Q84NG7|Q84NG7_HORVU      -------------------------------------------

 

11S GLOBULIN

Scores Table

SeqA Name                     Len(aa)  SeqB Name                     Len(aa)  Score

===================================================================================

1    tr|Q9AUD2|Q9AUD2_SESIN   497      2    sp|P19084|11S3_HELAN     493      34  

1    tr|Q9AUD2|Q9AUD2_SESIN   497      3    tr|Q2TLW0|Q2TLW0_SINAL   510      37  

1    tr|Q9AUD2|Q9AUD2_SESIN   497      4    tr|O23211|O23211_ARATH   499      13  

1    tr|Q9AUD2|Q9AUD2_SESIN   497      5    tr|O65042|O65042_ORYSA   174      44  

1    tr|Q9AUD2|Q9AUD2_SESIN   497      6    tr|Q84NG7|Q84NG7_HORVU   224      12  

1    tr|Q9AUD2|Q9AUD2_SESIN   497      7    tr|Q38712|Q38712_AMAHP   501      43  

1    tr|Q9AUD2|Q9AUD2_SESIN   497      8    tr|O82437|O82437_COFAR   490      48  

1    tr|Q9AUD2|Q9AUD2_SESIN   497      9    tr|A1E2B1|A1E2B1_ARAHY   277      32  

2    sp|P19084|11S3_HELAN     493      3    tr|Q2TLW0|Q2TLW0_SINAL   510      36  

2    sp|P19084|11S3_HELAN     493      4    tr|O23211|O23211_ARATH   499      10  

2    sp|P19084|11S3_HELAN     493      5    tr|O65042|O65042_ORYSA   174      40  

2    sp|P19084|11S3_HELAN     493      6    tr|Q84NG7|Q84NG7_HORVU   224      12  

2    sp|P19084|11S3_HELAN     493      7    tr|Q38712|Q38712_AMAHP   501      41  

2    sp|P19084|11S3_HELAN     493      8    tr|O82437|O82437_COFAR   490      40  

2    sp|P19084|11S3_HELAN     493      9    tr|A1E2B1|A1E2B1_ARAHY   277      26  

3    tr|Q2TLW0|Q2TLW0_SINAL   510      4    tr|O23211|O23211_ARATH   499      11  

3    tr|Q2TLW0|Q2TLW0_SINAL   510      5    tr|O65042|O65042_ORYSA   174      38  

3    tr|Q2TLW0|Q2TLW0_SINAL   510      6    tr|Q84NG7|Q84NG7_HORVU   224      8   

3    tr|Q2TLW0|Q2TLW0_SINAL   510      7    tr|Q38712|Q38712_AMAHP   501      44  

3    tr|Q2TLW0|Q2TLW0_SINAL   510      8    tr|O82437|O82437_COFAR   490      40  

3    tr|Q2TLW0|Q2TLW0_SINAL   510      9    tr|A1E2B1|A1E2B1_ARAHY   277      27  

4    tr|O23211|O23211_ARATH   499      5    tr|O65042|O65042_ORYSA   174      13  

4    tr|O23211|O23211_ARATH   499      6    tr|Q84NG7|Q84NG7_HORVU   224      8   

4    tr|O23211|O23211_ARATH   499      7    tr|Q38712|Q38712_AMAHP   501      15  

4    tr|O23211|O23211_ARATH   499      8    tr|O82437|O82437_COFAR   490      15  

4    tr|O23211|O23211_ARATH   499      9    tr|A1E2B1|A1E2B1_ARAHY   277      11  

5    tr|O65042|O65042_ORYSA   174      6    tr|Q84NG7|Q84NG7_HORVU   224      4   

5    tr|O65042|O65042_ORYSA   174      7    tr|Q38712|Q38712_AMAHP   501      44  

5    tr|O65042|O65042_ORYSA   174      8    tr|O82437|O82437_COFAR   490      39  

5    tr|O65042|O65042_ORYSA   174      9    tr|A1E2B1|A1E2B1_ARAHY   277      10  

6    tr|Q84NG7|Q84NG7_HORVU   224      7    tr|Q38712|Q38712_AMAHP   501      12  

6    tr|Q84NG7|Q84NG7_HORVU   224      8    tr|O82437|O82437_COFAR   490      13  

6    tr|Q84NG7|Q84NG7_HORVU   224      9    tr|A1E2B1|A1E2B1_ARAHY   277      5   

7    tr|Q38712|Q38712_AMAHP   501      8    tr|O82437|O82437_COFAR   490      41  

7    tr|Q38712|Q38712_AMAHP   501      9    tr|A1E2B1|A1E2B1_ARAHY   277      31  

8    tr|O82437|O82437_COFAR   490      9    tr|A1E2B1|A1E2B1_ARAHY   277      29  

===================================================================================

 

 

CLUSTAL 2.0.12 multiple sequence alignment

sp|P19084|11S3_HELAN        --------------MASKATLLLA---FTLLFATCIAR------------ 21

tr|O65042|O65042_ORYSA      --------------------------------------------------

tr|Q2TLW0|Q2TLW0_SINAL      --------------MVKLAHLLVATVGVLLVLNGCLARQS---------- 26

tr|Q38712|Q38712_AMAHP      STHASGFFFFHPTKMAKSTNYFLISCLLFVLFNGCMGEGR---------- 40

tr|Q9AUD2|Q9AUD2_SESIN      ---------------MALTSLLSFFIVVTLLIRGLSAQLAGEQDFYWQDL 35

tr|O82437|O82437_COFAR      ---------------MAHSHMISLSLYVLLFL-GCLAQLG---------- 24

sp|Q9XHP0|11S2_SESIN        ----------------MVAFKFLLALSLSLLVSAAIAQTR---------- 24

tr|O23211|O23211_ARATH      ---------------MTRFAVLPLSVLLLVLLFLCTESLAKSEE------ 29

tr|Q84NG7|Q84NG7_HORVU      --------------------------------------------------

                                                                             

sp|P19084|11S3_HELAN        ---HQQRQQQQNQCQLQN---IEALEPIEVIQAEAGVTEIWDAYDQQFQC 65

tr|O65042|O65042_ORYSA      --------------------------------------------------

tr|Q2TLW0|Q2TLW0_SINAL      ---LGVPPQVKDACNLDN---LDVLQPTEVIKSEAGQVEYWDHNHPQIRC 70

tr|Q38712|Q38712_AMAHP      ---FREFQQG-NECQIDR---LTALEPTNRIQAERGLTEVWDSNEQEFRC 83

tr|Q9AUD2|Q9AUD2_SESIN      QSQQQHKLQARTDCRVER---LTAQEPTIRFESEAGLTEFWDRNNQQFEC 82

tr|O82437|O82437_COFAR      --RPQPRLRGKTQCDIQK---LNAQEPSFRFPSEAGLTEFWDSNNPEFGC 69

sp|Q9XHP0|11S2_SESIN        ----EPRLTQGQQCRFQR---ISGAQPSLRIQSEGGTTELWDERQEQFQC 67

tr|O23211|O23211_ARATH      --SEEYDVAVPSCCGFSSPLLIKKDQWKPIFETKFGQISTVQIGNGCGGM 77

tr|Q84NG7|Q84NG7_HORVU      --------------------------------------------------

 sp|P19084|11S3_HELAN        A-----WSILFDTGFNLVAFSCLPTSTPLFWPSSREGVILPGCRRTYEY- 109

tr|O65042|O65042_ORYSA      --------------------------------------------------

tr|Q2TLW0|Q2TLW0_SINAL      AGVSIARLVIQKGGLYLPTFFSSPFISYVVQGMGISGRVIPGCAETFMDS 120

tr|Q38712|Q38712_AMAHP      AGVSVIRRTIEPHGLLLPSFTSAPELIYIEQGNGITGMMIPGCPETYES- 132

tr|Q9AUD2|Q9AUD2_SESIN      AGVAAVRNVIQPRGLLLPHYNNAPQLLYVVRGRGIQGTVIPGCAETFER- 131

tr|O82437|O82437_COFAR      AGVEFERNTVQPKGLRLPHYSNVPKFVYVVEGTGVQGTVIPGCAETFESQ 119

sp|Q9XHP0|11S2_SESIN        AGIVAMRSTIRPNGLSLPNYHPSPRLVYIERGQGLISIMVPGCAETYQVH 117

tr|O23211|O23211_ARATH      GPYKIHSITLEPNTILLPLLLHSDMVFFVDSGSGILNWVDEEAKSTEIRL 127

tr|Q84NG7|Q84NG7_HORVU      --------------------------------------------------

sp|P19084|11S3_HELAN        -------SQEQQFSGEGGRRGGGEGT------------------------ 128

tr|O65042|O65042_ORYSA      --------------------------------------------------

tr|Q2TLW0|Q2TLW0_SINAL      QPMQGQQGQQQGQQGQQGQQGQQGQQGQQGQQGQQGQQGQQGQQGQQGQQ 170

tr|Q38712|Q38712_AMAHP      -------GSQQFQGGEDERIREQGSR-------------------KFGMR 156

tr|Q9AUD2|Q9AUD2_SESIN      -------DTQP--------------RQDRR-------------------- 140

tr|O82437|O82437_COFAR      GESFWGGQEQPGKGQEGQEQGSKGGQEGRR-------------------- 149

sp|Q9XHP0|11S2_SESIN        R------SQRTMERTEASEQQDRG-------------------------- 135

tr|O23211|O23211_ARATH      GDVYRLRPGSVFYLQSKPDPCFGAYS------------------------ 153

tr|Q84NG7|Q84NG7_HORVU      --------------------------------------------------

 sp|P19084|11S3_HELAN        ---FRTVIRKLENLKEGDVVAIPTGTAHWLHNDGNT-ELVVVFLDTQNHE 174

tr|O65042|O65042_ORYSA      --------------------------------------------------

tr|Q2TLW0|Q2TLW0_SINAL      GQGFRDMHQKVEHVRHGDAIAMTPGSAQWIYNTGDQPLIIVSLIDIANYQ 220

tr|Q38712|Q38712_AMAHP      GDRFQDQHQKIRHLREGDIFAMPAGVSHWAYNNGDQPLVAVILIDTANHA 206

tr|Q9AUD2|Q9AUD2_SESIN      -RRFMDRHQKVRQFRQGDILALPAGLTLWFYNNGGEPLITVALLDTGNAA 189

tr|O82437|O82437_COFAR      -QRFPDRHQKLRRFQKGDVLILLPGFTQWTYNDGDVPLVTVALLDVANEA 198

sp|Q9XHP0|11S2_SESIN        --SVRDLHQKVHRLRQGDIVAIPSGAAHWCYNDGSEDLVAVSINDVNHLS 183

tr|O23211|O23211_ARATH      --SITDLMFGFDETILQSAFGVPEGIIELMRNRTKPPLIVSETLCTPGVA 201

tr|Q84NG7|Q84NG7_HORVU      --------------------------------MGKFIFFAVFLTTLVTIS 18

                                                                             

sp|P19084|11S3_HELAN        NQLDEN-QRRFFLAGNPQAQAQSQQQQQRQPRQQSPQRQRQRQRQGQGQN 223

tr|O65042|O65042_ORYSA      ------------------------------------------ETERGGEE 8

tr|Q2TLW0|Q2TLW0_SINAL      NQLDRN-PRTFRLAGNNQ-------------QGSSQQQQQQQQ------- 249

tr|Q38712|Q38712_AMAHP      NQLDKNFPTRFYLAGKPQ-------------QEHSGEHQFSRESRRGERN 243

tr|Q9AUD2|Q9AUD2_SESIN      NQLDQT-FRHFFLAGNPQ--------------GGRQSYFGRPQTEKQQGE 224

tr|O82437|O82437_COFAR      NQLDLQ-SRKFFLAGNPQ--------------QGG----GKEGHQGQQQQ 229

sp|Q9XHP0|11S2_SESIN        NQLDQK-FRAFYLAGGVP--------------------RSGEQEQQARQT 212

tr|O23211|O23211_ARATH      NTWQLQPRLLKLFAGSAD--------------LVDNKKKKEKKEKKEKVK 237

tr|Q84NG7|Q84NG7_HORVU      --------------------------------------------------

                                                                             

sp|P19084|11S3_HELAN        AGNIFNGFTPELIAQSFNVDQETAQKLQGQNDQRGHIVNVG-QDLQIVRP 272

tr|O65042|O65042_ORYSA      SYNILSGFDTELLAESMRVSPDIARKLQGRSDKRGNIVRVRRGGLRMLRP 58

tr|Q2TLW0|Q2TLW0_SINAL      --NILSGFDPQVLAQALKIDVRLAQELQNQQDKRGNIVRVK-GPFQVVRP 296

tr|Q38712|Q38712_AMAHP      TGNIFRGFETRLLAESFGVSEEIAQKLQAEQDDRGNIVRVQ-EGLHVIKP 292

tr|Q9AUD2|Q9AUD2_SESIN      TKNIFNGFDDEILADAFGVDVQTARRLKGQDDLRGRIVRAE--RLDIVLP 272

tr|O82437|O82437_COFAR      HRNIFSGFDDQLLADAFNVDLKIIQKLKGPKDQRGSTVRAE--KLQLFLP 277

sp|Q9XHP0|11S2_SESIN        FHNIFRAFDAELLSEAFNVPQETIRRMQSEEEERGLIVMAR-ERMTFVRP 261

tr|O23211|O23211_ARATH      KAKTFNVFESEPDFESP-YGRTITINRKDLKVLKGSMVGVS---MVNLTQ 283

tr|Q84NG7|Q84NG7_HORVU      ---AAQGVLEQSLADAQCRGEVQAKPLLACRQILEHQLTGR---AVGVRP 62

                                   .  .   ::                     :         . 

sp|P19084|11S3_HELAN        PQDRRSPRQQQEQATSPRQQQEQQQGRRGGWSNGVEETICSMKFKVNIDN 322

tr|O65042|O65042_ORYSA      ATERVTDEEMMRGANAAAAAG-----------NGIDEAVCLMKLRENVAD 97

tr|Q2TLW0|Q2TLW0_SINAL      PLRQAYESEQWRHPRGPPQSPQD---------NGLEETICSMRTHENIDD 337

tr|Q38712|Q38712_AMAHP      PSRAWEEREQG--SRGSRYLP-----------NGVEETICSARLAVNVDD 329

tr|Q9AUD2|Q9AUD2_SESIN      GEEEEER-----WERDPYSG-----------ANGLEETLCTAKLRENLDE 306

tr|O82437|O82437_COFAR      EYSEQEQQPQQQQEQQQHGVGRGWR------SNGLEETLCTVKLSENIGL 321

sp|Q9XHP0|11S2_SESIN        DEEEGEQ------EHRGRQLD-----------NGLEETFCTMKFRTNVES 294

tr|O23211|O23211_ARATH      GSMMGPHWNPWACEISIVLKG-----------AGMVRVLRSSISSNTSSE 322

tr|Q84NG7|Q84NG7_HORVU      FQAQWGARDRCCQQLESVSRG------------------CRCSALRGMVR 94

                                                                             

sp|P19084|11S3_HELAN        PSQADFVNPQAGSIANLNSFKFPILEHLRLSVERGELRPNAIQSPHWTIN 372

tr|O65042|O65042_ORYSA      PMKADLYTPNGGRITVLNSQKLPVLKLIKMSVNRGVMRRNAILAPHWNIN 147

tr|Q2TLW0|Q2TLW0_SINAL      PARADIYKPNLGRVTSVNSYTLPILQYIRLSATRGILQGSAMVLPKYNMN 387

tr|Q38712|Q38712_AMAHP      PSKADVYTPEAGRLTTVNSFNLPILRHLRLSAAKGVLYRNAMMAPHYNLN 379

tr|Q9AUD2|Q9AUD2_SESIN      PARADVYNPHGGRISSLNSLTLPVLSWLRLSAEKGVLYRNGLVAPHWNLN 356

tr|O82437|O82437_COFAR      PQEADVFNPRAGRITTVNSQKIPILSSLQLSAERGFLYSNAIFAPHWNIN 371

sp|Q9XHP0|11S2_SESIN        RREADIFSRQAGRVHVVDRNKLPILKYMDLSAEKGNLYSNALVSPDWSMT 344

tr|O23211|O23211_ARATH      CKNVRFKVEEGDIFAVPRLHPMAQMSFNNDSLVFVGFTTSAKNNEPQFLA 372

tr|Q84NG7|Q84NG7_HORVU      DYEQSMPPLREGRRRSSGE-----------RQQEQGCSGESTAEQQQEVQ 133

                              .  .   . .                           ..       :

 

sp|P19084|11S3_HELAN        AHNLLYVTEGALRVQIVDNQGNSVFDNELREGQVVVIPQNFAVIKRAN-E 421

tr|O65042|O65042_ORYSA      AHAAVYATSGSAKLQVVSNEGRPVFDG----------------------- 174

tr|Q2TLW0|Q2TLW0_SINAL      ANEILYCTQGQARIQVVNDNGQNVLDQQVQKGQLVVIPQGFAYVVQSQ-N 436

tr|Q38712|Q38712_AMAHP      AHNIMYCVRGRGRIQIVNDQGQSVFDEELSRGQLVVVPQNFAIVKQAF-E 428

tr|Q9AUD2|Q9AUD2_SESIN      AHSIIYITRGSGRFQVVGHTGRSVFDGVVREGQLIIVPQNYVVAKRASQD 406

tr|O82437|O82437_COFAR      AHSALYVIRGNARIQVVDHKGNKVFDDEVKQGQLIIVPQYFAVIKKAG-N 420

sp|Q9XHP0|11S2_SESIN        GHTIVYVTRGDAQVQVVDHNGQALMNDRVNQGEMFVVPQYYTSTARAG-N 393

tr|O23211|O23211_ARATH      GEDSALRMLDRQVLAASLNVSSVTIDGLLGAQKEAVILECHSCAEGEIEK 422

tr|Q84NG7|Q84NG7_HORVU      GGQYGSETGESQQQQGGGYHGVTVGRGGQQQGQMLCRERPQRQQQGEG-- 181

                            .                   .                            

sp|P19084|11S3_HELAN        QGSRWVSFKTNDNAMIANLAGRVSASAASPLTLWANRYQLSREEAQQLKF 471

tr|O65042|O65042_ORYSA      --------------------------------------------------

tr|Q2TLW0|Q2TLW0_SINAL      N-FEWISFKTNANAMISTLAGRTSALRALPLEVITNAYQISLEEARKIKF 485

tr|Q38712|Q38712_AMAHP      DGFEWVSFKTSENAMFQSLAGRTSAIRSLPIDVVSNIYQISREEAFGLKF 478

tr|Q9AUD2|Q9AUD2_SESIN      EGLEWISFKTNDNAMTSQLAGRLSAIRAMPEEVVMTAYQVSRDEARRLKY 456

tr|O82437|O82437_COFAR      QGFEYVAFKTNDNAMINPLVGRLSAFRAIPEEVLRSSFQISSEEAEELKY 470

sp|Q9XHP0|11S2_SESIN        NGFEWVAFKTTGSPMRSPLAGYTSVIRAMPLQVITNSYQISPNQAQALKM 443

tr|O23211|O23211_ARATH      LKVEIERKKIDDERKRRHDERKKEEEEAKREEEERRKREEEEEKKRWPPQ 472

tr|Q84NG7|Q84NG7_HORVU      --------FSGEGAQQKPKVGRVRLTKVR----LPTACRIEPQECSVFST 219

                                                                             

sp|P19084|11S3_HELAN        SQRETVLFAPSFSRGQGIRASR------------------- 493

tr|O65042|O65042_ORYSA      -----------------------------------------

tr|Q2TLW0|Q2TLW0_SINAL      NTLETTLTRARGGQQPQLIEEIVEV---------------- 510

tr|Q38712|Q38712_AMAHP      NRPETTLFRSSGQGEYRRKISIA------------------ 501

tr|Q9AUD2|Q9AUD2_SESIN      NREESRVFSSTSRYSWPRSSRPMSYMPKPFEYVLDVIKSMM 497

tr|O82437|O82437_COFAR      GRQEALLLSEQS----QQGKREVA----------------- 490

sp|Q9XHP0|11S2_SESIN        NRGSQSFLLSPGGRRS------------------------- 459

tr|O23211|O23211_ARATH      QPPQEEELRERQLPMEKEWEMEGEEES-------------- 499

tr|Q84NG7|Q84NG7_HORVU      LPVLG------------------------------------ 224

 

 

 

PROTEIN SEQUENCE ANALYSIS:

The protein sequences for 7S globulin and 11S globulin were obtained from Uniprot with Accession numbers Q9AUD2-SESIN and Q9AUDO-SESIN respectively.  Similarity studies were carried out using BLAST and Clustal W tools.

 

SIMILARITY STUDIES USING BLAST:

 In BLAST result, 111 hits were obtained for 7S globulin and 100 hits for 11S globulin.  The bit score 1201(E.score=e-0.0) was found for the query sequence 7S globulin and the next bit score 514 was found for Corylus avellana-glycoprotein precursor (E.score=1e-143) Minimum bit score 243 (E. score=6e-62) was obtained for Soyabean Beta Conglycinine Beta Homotrimer. The bit score 882(E.score=e-0.0) was found for query sequence 11S globulin and the next bit scores 491, 468 was found for 11S globulin precursor isoform of Sesamum indicum (E.score =7e-137), 11S globulin like protein of Actinidia chinensis (8e-130) respectively. Minimum bit score 358(E.score=8e-97) was for Oryza sativa Japonica glutelin.

 

MULTIPLE SEQUENCE ALIGNMENT:

In multiple sequence alignment with ClustalW, 7S globulin of Sesamum indicum (Q9AUDO-SESIN)was compared with basic 7S globulin of  Glycinemax (7SBI-SOYBN)Zamia furfuracea (Q41727-ZAMFU), Hordeum vulgare (Q84NG7-HORVU)7 Elaeis guineensis var tenera (Q9AU64-ELAGV),Globulin1 of  Oryza sativa (065042-ORYSA)and globulin like protein of Arabidopsis thaliana(0232H-ARATH).7S globulin of Sesamum indicum shows more similarity with 7S globulin of Elaeis guineensis var tenera (Q9AU64-ELAGV:score =42).The results also show that  Zamia furfuracea has a score of  32 , Hordeum vulgare with 13, Glycinemax having score 4, Oryza sativa with 18, Arabidopsis thaliana with 16 while aligning with 7S globulin of Sesamum indicum.

 

Similarily 11S globulin of Sesamum indicum(Q9AUD2-SESIN)was compared with 11S globulin of Sinapsis alba(Q2DLWO-SINAL),Globulin like protein of Arabidopsis thaliana (023211-ARATH), Oryza sativa(065042-ORYSA), Hordeum vulgare(Q84NG7-HORVU),11S globulin seedstorage protein of  Helianthus annus(11S3-HELAN), Amaranthus hypochondriachus (Q38712-AMAHR), Coffea arabica(082437-COFAR)and Arachis hypogaea (AIE2B1-ARAHY). 11S globulin of Sesamum indicum shows more similarity with 11S storage globulin of Coffea arabica(082437-COFAR-score=48). The results also show that    Sinapsis alba has a score of  37 , Hordeum vulgare with 12, Helianthus annus having score 34, Oryza sativa with 44, Arabidopsis thaliana with 13, Amaranthus hypochondriachus with 43, Arachis hypogaea  having score of 32 while aligning with 11S globulin of Sesamum indicum.

 

 

 

STRUCTURAL ANALYSIS

PRIMARY STRUCTURE ANALYSIS (Protparam)

7S GLOBULIN

Number of amino acids: 585 Molecular weight: 67068.9 Theoretical pI: 7.55
Amino acid composition:

Top of Form

Ala (A)  37      6.3%

Arg (R)  62    10.6%

Asn (N)  23      3.9%

Asp (D)  19      3.2%

Cys (C)  15      2.6%

Gln (Q)  54      9.2%

Glu (E)  67    11.5%

Gly (G)  52      8.9%

His (H)  14      2.4%

Ile (I)  25      4.3%

Leu (L)  38      6.5%

Lys (K)  25      4.3%

Met (M)   9      1.5%

Phe (F)  25      4.3%

Pro (P)  20      3.4%

Ser (S)  32      5.5%

Thr (T)  19      3.2%

Trp (W)   3      0.5%

Tyr (Y)  15      2.6%

Val (V)  31      5.3%

Pyl (O)   0      0.0%

Sec (U)   0      0.0%

Bottom of Form

Total number of negatively charged residues (Asp + Glu): 86

Total number of positively charged residues (Arg + Lys): 87

 

Atomic composition:

Carbon      C        2882

Hydrogen    H        4570

Nitrogen    N         904

Oxygen      O         901

Sulfur              S                    24
 
 
 
 
11S GLOBULIN
Number of amino acids: 497 Molecular weight: 56587.9 Theoretical pI: 8.57
Amino acid composition
Ala (A)  36      7.2%
Arg (R)  50     10.1%
Asn (N)  23      4.6%
Asp (D)  25      5.0%
Cys (C)   4      0.8%
Gln (Q)  34      6.8%
Glu (E)  34      6.8%
Gly (G)  38      7.6%
His (H)   8      1.6%
Ile (I)  21      4.2%
Leu (L)  49      9.9%
Lys (K)  12      2.4%
Met (M)   9      1.8%
Phe (F)  22      4.4%
Pro (P)  22      4.4%
Ser (S)  29      5.8%
Thr (T)  25      5.0%
Trp (W)   8      1.6%
Tyr (Y)  15      3.0%
Val (V)  33      6.6%
Pyl (O)   0      0.0%
Sec (U)   0      0.0%
Total number of negatively charged residues (Asp + Glu): 59
Total number of positively charged residues (Arg + Lys): 62
Atomic composition:
Carbon      C        2496
Hydrogen    H        3924
Nitrogen    N         740
Oxygen      O         742
Sulfur      S          13
 
SECONDARY STRUCTURE PREDICTION
7S GLOBULIN
10        20        30        40        50        60        70
         |         |         |         |         |         |         |
MSCGGRLCLVLFALLLASAVVASESKDPELKQCKHQCKAQQQISKEQKEACIQACKEYIRQKHQGEHGRG
ccccccchhhhhhhhhhhhhhhhhccchhhhhhcccccccchhhhhchhhhhhhhhhhhhhccccccccc
GGDILEEEVWNRKSPIERLRECSRGCEQQHGEQREECLRRCQEEYQREKGRQDDDNPTDPEKQYQQCRLQ
ccchhhhhhhccccceeeeeeccccccccccccchhhhhhchhhhhhhcccccccccccchhhhhhheee
CRRQGEGGGFSREHCERRREEKYREQQGREGGRGEMYEGREREEEQEEQGRGRIPYVFEDQHFITGFRTQ
eeccccccccchhhhhhchhhhhhhhhcccccccccccchhhhhhhhhhcccccceeccccceeeeeeee
HGRMRVLQKFTDRSELLRGIENYRVAILEAEPQTFIVPNHWDAESVVFVAKGRGTISLVRQDRRESLNIK
ccchhhhhhccchhhhhhhhhhhhhhhhhcccceeeeccccccceeeeecccccceeehhhhhhhhhhhh
QGDILKINAGTTAYLINRDNNERLVLAKLLQPVSTPGEFELFFGAGGENPESFFKSFSDEILEAAFNTRR
ccceeeeccccceeeeccccchhhhhhhhccccccccceeeeecccccccccccccchhhhhhhhhhhhh
DRLQRIFGQQRQGVIVKASEEQVRAMSRHEEGGIWPFGGESKGTINIYQQRPTHSNQYGQLHEVDASQYR
hhhhhhhhhhcchhhhhhhhhhhhhhhhhcccceeeccccccceeeeeeccccccccccchhhhhhhhhh
QLRDLDLTVSLANITQGAMTAPHYNSKATKIALVVDGEGYFEMACPHMSRSRGSYQGETRGRPSYQRVAS
hhhhhhhhhhhhhhccccccccccccccceeeeeeccccceeeeccccccccccccccccccccchhhhc
RLTRGTVVIIPAGHPFVAVASSNQNLQVLCFEVNANNNEKFPLAGRRNVMNQLEREAKELAFGMPAREVE
cccccceeeecccccceeeeccccceeeeeeeecccccccccccchhhhhhhhhhhhhhhhhcccchhhh
EVSRSQQEEFFFKGPRQQQQGRADA
hhhhhhhhhhhcccchhhhceeeec
Sequence length :   585
GOR4 :
   Alpha helix     (Hh) :   241 is  41.20%
   310  helix       (Gg) :     0 is   0.00%
   Pi helix        (Ii) :     0 is   0.00%
   Beta bridge     (Bb) :     0 is   0.00%
   Extended strand (Ee) :    85 is  14.53%
   Beta turn       (Tt) :     0 is   0.00%
   Bend region     (Ss) :     0 is   0.00%
   Random coil     (Cc) :   259 is  44.27%
   Ambigous states (?)  :     0 is   0.00%
 
Other states :0 is 0.00%
 

11S GLOBULIN

10        20        30        40        50        60        70
         |         |         |         |         |         |         |
MALTSLLSFFIVVTLLIRGLSAQLAGEQDFYWQDLQSQQQHKLQARTDCRVERLTAQEPTIRFESEAGLT
cccccccchhhhhhhhhhchhhhhhhhhchhhhhhhhhhhhhhcccccccccccccccchhhhhhhccch
EFWDRNNQQFECAGVAAVRNVIQPRGLLLPHYNNAPQLLYVVRGRGIQGTVIPGCAETFERDTQPRQDRR
hhhccccceeccccchhhhcccccccceecccccccceeeeeccccccceeecccccccccccccchhhh
RRFMDRHQKVRQFRQGDILALPAGLTLWFYNNGGEPLITVALLDTGNAANQLDQTFRHFFLAGNPQGGRQ
hhhhhhhhhhhhhhhchhhhccccceeeeecccccceeeeeeeeeccccchhhhhhhheeeccccccccc
SYFGRPQTEKQQGETKNIFNGFDDEILADAFGVDVQTARRLKGQDDLRGRIVRAERLDIVLPGEEEEERW
cccccccchhhhcccceeccccchhhhhhhccchhhhhhhhhcccchhhhhhhhhhhceeccccchhhhh
ERDPYSGANGLEETLCTAKLRENLDEPARADVYNPHGGRISSLNSLTLPVLSWLRLSAEKGVLYRNGLVA
cccccccccchhhhhhhhhhhccccccccccccccccceeeeccccccchhhhhhhhhhhceeeecceee
PHWNLNAHSIIYITRGSGRFQVVGHTGRSVFDGVVREGQLIIVPQNYVVAKRASQDEGLEWISFKTNDNA
cccccccceeeeeeccccceeeeccccceeeceeccccceeeeccchhhhhhhccccceeeeeeecccch
MTSQLAGRLSAIRAMPEEVVMTAYQVSRDEARRLKYNREESRVFSSTSRYSWPRSSRPMSYMPKPFEYVL
hhhhhhhhhhhhhccchhhhhhhhhhhhhhhhhhhhccccceeeeecccccccceccccccccccceeee
DVIKSMM
eeeecee
 
 
 
 
Sequence length :   497
GOR4 :
   Alpha helix     (Hh) :   178 is  35.81%
   310  helix       (Gg) :     0 is   0.00%
   Pi helix        (Ii) :     0 is   0.00%
   Beta bridge     (Bb) :     0 is   0.00%
   Extended strand (Ee) :    86 is  17.30%
   Beta turn       (Tt) :     0 is   0.00%
   Bend region     (Ss) :     0 is   0.00%
   Random coil     (Cc) :   233 is  46.88%
   Ambigous states (?)  :     0 is   0.00%
   Other states         :     0 is   0.00%

SECONDARY STRUCTURE PREDICTION

Secondary structure predicted using GOR (Garnier, Osguthorpe and Robson) method shows that 7S globulin has 41.20% alpha helix, 14.53% extended strand and 44.27% random coil .In case of 11S globulin, there are 35.81% alpha helix, 17.30%,extended strand and 46.88% random coil. These results clearly reveal the presence of alpha helix, extended strand and random coils in7S and 11S globulin with a high occurrence of random coils in both the globulin sub units.

 

CONCLUSION:

The present study conducted with the hexane and methanolic extract of Sesamum indicum reveals that it can contribute to useful amounts of aminoacids to human diet. The quantitative analysis of proteins in Sesamum indicum seeds reveal that sesame seeds are one of the good sources of proteins and the proteins are presents at levels sufficient enough to benefit health. So, it can be supplemented for malnourished people. The insilico analysis of protein shows that the sequences of 7Sglobulin and 11S globulin of Sesamum indicum have greater sequence homology with 7S globulin of Elaeis guineensis var tenera and 11S storage globulin of Coffea arabica respectively. The current study clearly reveals the presence of alpha helix, extended strand and random coils in7S and 11S globulin with a high occurrence of random coils in both the globulin subunits. Further analysis can be performed to analyze the conserved sequences of these proteins and derive the secondary and tertiary structures of these globulins.

 

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Received on 08.12.2016       Accepted on 08.01.2017     

© Asian Pharma Press All Right Reserved

Asian J. Pharm. Tech.  2017; 7(1): 37-50.

DOI: 10.5958/2231-5713.2017.00007.1

 

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