Patent Application: US-81709097-A

Abstract:
the present invention concerns a new phosphatidyl - inositol - 3 - kinase , a nucleic acid coding for it , an antibody directed against the protein as well as the diagnostic and therapeutic use of the protein , the nucleic acid and the antibody .

Description:
seq id no . 1 shows a nucleic acid sequence which contains genetic information coding for pi3kγ and seq id no . 2 shows the amino acid sequence of a pi3kγ . seq id no : 3 shows a nucleic acid sequence which contains genetic information coding for a further pi3kγ and seq id no . 4 shows the amino acid sequence of a further pi3kγ . in order to isolate cdna sequences which code for new pi3k a human bone marrow cdna library was screened using the polymerase chain reaction ( pcr ). for this degenerate oligonucleotide primers were used corresponding to the amino acid sequences kngddlr seq id no : 6 ! and hidfg seq id no : 7 !. a 402 bp long fragment was obtained . this fragment was subcloned and sequenced . several overlapping clones from a human u937 cdna library were isolated using this pcr fragment as a probe . the largest clone contained the nucleic acid sequence shown in seq id no . 1 with an open reading frame which codes for a protein with 1049 amino acids ( seq id no . 2 ). this protein named pi3kγ has a molecular mass of approximately 120 kda . a further pi3kγ sequence which was obtained from a cdna library is shown in seq id no . 3 and codes for a protein with 1050 amino acids ( seq id no . 4 ). a polyclonal rabbit antiserum against pi3kγ was produced by immunization with a 15 amino acid long peptide of the sequence nsqlpesfrvpydpg seq id no : 5 ( corresponding to amino acids 741 to 755 in seq id no . 2 or amino acids 742 to 756 in seq id no . 4 ). the serum was purified by protein a chromatography and affinity chromatography using the peptide antigen coupled to actigel ( sterogene ). pi3kγ can be detected as a 110 kda protein using this antiserum by immunoprecipitation and western blot analysis of u937 and k562 cells ( fig1 ). the immunoprecipitation and western blot were carried out according to the method of hiles et al . ( cell 70 ( 1992 ) 419 - 429 ). a conjugate of horseradish peroxidase and anti - rabbit antiserum ( sigma , 1 : 2000 dilution ) was used as the secondary antibody . the bound peroxidase was visualized by chemiluminescence . a northern blot analysis of human tissue from the pancreas , kidney , skeletal muscle , liver , lung , placenta , brain and heart each showed different concentrations of a 5 . 3 kb long mrna . the dna coding for pi3kγ was cloned into the vector pacg2t ( davis et al ., biotechnology 11 ( 1993 ), 933 - 936 ). the resulting recombinant vector pacg2t - pi3kγ contained the pi3kγ cdna from codon 4 onwards in a fusion with the glutathione s transferase ( gst ) gene . sf9 cells were cotransfected with pacg2t - pi3kγ and linearized baculovirus dna ( baculogold , pharmingen ). individual recombinant baculovirus gst - pi3kγ plaques were purified and amplified . the expression and purification of recombinant protein were carried out using standard protocols ( dhand et al ., embo j . 13 ( 1994 ), 511 - 521 ). fig2 a shows the expression of recombinant pi3kγ gst fusion protein or gst alone after fractionation on a sds polyacrylamide gel . the detection was carried out by staining with coomassie blue . fig2 b shows a detection of the enzymatic activity of purified recombinant pi3ky . phosphatidylinositol ( pi , lane 1 ), phosphatidylinositol - 4 - phosphate ( pi4 - p , lane 2 ) and phosphatidylinositol - 4 , 5 - diphosphate ( pi 4 , 5 - p 2 , lane 3 ) were used as substrates . the test was carried out essentially according to the method of stephens et al . ( cell 77 ( 1994 ), 83 - 93 ) but without cholate . 30 μl sonicated lipid vesicles containing 320 μm phosphatidyl ethanolamine , 140 μm phosphatidyl choline , 300 μm phosphatidyl serine , 30 μm sphingomyelin and 320 μm substrate were added to 10 μl enzyme ( 0 . 1 ng ) and incubated for 8 minutes on ice . the test was started by addition of 10 μl 20 μm atp containing 10 μci γ ( 32 p ) atp and incubated for 15 minutes at room temperature . the extracted lipids were separated and visualized . the identity of the 3 - phosphorylated phosphoinositides was confirmed by anion exchange hplc after deacylation of the lipids ( auger et al ., cell 57 ( 1989 ), 167 - 175 ). fig2 b shows that the substrates were phosphorylated in the d - 3 position of the inositol ring . in addition it was found that pi3kγ was inhibited by wortmannin at nanomolar concentrations . in order to detect an interaction between the regulatory and adaptory subunits of p85 ( p85α and p85β ) and pi3kγ , human pi3kγ and bovine pi3kα ( hiles et al ., supra ) were expressed in sf9 insect cells according to the process described in example 3 as gst fusion proteins either alone or together with p85α and p85β . after lysis and centrifugation of the cells , glutathione - sepharose was added to the supernatant in order to bind the gst fusion proteins . the particles were washed and analysed by sds - page and western - blot . pi3kγ was detected using the polyclonal antipeptide antibody described in example 2 . appropriate mixtures of specific monoclonal antibodies ( dhand et al ., supra ) as a primary antibody and conjugates of horseradish peroxidase and anti - mouse antibodies ( dianova , 1 : 4000 dilution ) as the secondary antibody were used to detect p85α , p85β and bovine pi3kα . fig3 shows that pi3kγ in contrast to pi3kα does not bind to p85α and p85β subunits . the regulation mechanism of pi3kγ is therefore different from pi3kα . in order to detect an interaction between pi3kγ and g proteins , purified recombinant human pi3kγ and bovine pi3kα was incubated with the transducin species g t βγ . a substantial amplification of the kinase activity of pi3kγ was found . this stimulation was suppressed by adding gtp - loaded g t α which confirms the specificity of the interaction . in contrast the enzymatic activity of pi3kα could not be stimulated by addition of g t βγ . a similar activation of the enzyme was found after addition of gα in the presence of 20 μm alcl 3 and 10 mm naf . the complex gα - gdp - alf 4 - acts as an activating species . the determination of activity was carried out as described in example 3 . the g t proteins purified from bovine retina according to the method of camps et al . ( nature 360 ( 1992 ) 684 - 686 ) were incubated on ice with the lipid vesicles for 5 min ( g t βγ ) or for 20 min ( g t βγ plus g t α - gtp ) before adding the enzyme . binding studies with recombinant pi3kγ showed that the regulator protein ras in its active gtp - loaded form associates with the enzyme . ras in its constitutively active version is able to oncogenically transform cells . hence this is a further indication for the potential importance of pi3kγ in the regulation of cellular proliferation . __________________________________________________________________________sequence listing ( 1 ) general information :( iii ) number of sequences : 7 ( 2 ) information for seq id no : 1 :( i ) sequence characteristics :( a ) length : 4134 base pairs ( b ) type : nucleic acid ( c ) strandedness : both ( d ) topology : linear ( ii ) molecule type : cdna ( vi ) original source :( a ) organism : homo sapiens ( ix ) feature :( a ) name / key : cds ( b ) location : 423 .. 3569 ( xi ) sequence description : seq id no : 1 : gaattcggcacgagcacttccttctcggctagattatctgaaactgttgtcggttcttga60gatgatactaccaccgaatgtctgtgtttcattgtctagtccaacctgtattgtggatat120ctacaacgttccggcaatagttttgcaggtgcatcacatttttgtttttgttttgggagg180aaaagggagggcacggcagccaggcttcatattcctacaagtgcatgcttcaagattact240gtacttacagtgtttccaacatcttctcataaaaggggaaagcttcatagcctcaaccat300gaaggaaaccagtcgcatagggcatggagctggagaactataaacagcccgtggtgctga360gagaggacaactgccgaaggcgccggaggatgaagccgcgcagtgctgccagcctgtcct420ccatggagctcatccccatcgagttcgtgctgcccaccagccagcgc467metgluleuileproilegluphevalleuprothrserglnarg51015aaatgcaagagccccgaaacggcgctgctgcacgtggccggccacggc515lyscyslysserprogluthralaleuleuhisvalalaglyhisgly202530aacgtggagcagatgaaggcccaggtgtggctgcgagcgctggagacc563asnvalgluglnmetlysalaglnvaltrpleuargalaleugluthr354045agctggcgcggacttctaccaccggctgggaccgcatcacttcctcct611sertrpargglyleuleuproproalaglythralaserleupropro505560gctctatcagaagaagggcagtggtacgagatctacgacaagtaccag659alaleuserglugluglyglntrptyrgluiletyrasplystyrgln657075gtggtgcagactctggactgcctgcgctactggaaggccacgcaccgg707valvalglnthrleuaspcysleuargtyrtrplysalathrhisarg80859095agcccgggccagatccacctggtgcagcggcacccgccctccgaggag755serproglyglnilehisleuvalglnarghisproprosergluglu100105110tcccaagccttccagcggcagctcacggcgctgattggctatgacgtc803serglnalapheglnargglnleuthralaleuileglytyraspval115120125actgacgtcagcaacgtgcacgacgatgagctggagttcacgcgccgt851thraspvalserasnvalhisaspaspgluleugluphethrargarg130135140ggcttggtgaccccgcgcatggcggaggtggccagccgcgaccccaag899glyleuvalthrproargmetalagluvalalaserargaspprolys145150155ctctacgccatgcacccgtgggtgacgtccaagcccctcccggagtac947leutyralamethisprotrpvalthrserlysproleuproglutyr160165170175ctgtggaagaagattgccaacaactgcatcttcatcgtcattcaccgc995leutrplyslysilealaasnasncysilepheilevalilehisarg180185190agcaccaccagccagaccattaaggtctcacccgacgacacccccggc1043serthrthrserglnthrilelysvalserproaspaspthrprogly195200205gccatcctgcagagcttcttcaccaagatggccaagaagaaatctctg1091alaileleuglnserphephethrlysmetalalyslyslysserleu210215220atggatattcccgaaagccaaagcgaacaggattttgtgctgcgcgtc1139metaspileprogluserglnsergluglnaspphevalleuargval225230235tgtggccgggatgagtacctggtgggcgaaacgcccatcaaaaacttc1187cysglyargaspglutyrleuvalglygluthrproilelysasnphe240245250255cagtgggtgaggcactgcctcaagaacggagaagagattcacgtggta1235glntrpvalarghiscysleulysasnglyglugluilehisvalval260265270ctggacacgcctccagacccggccctagacgaggtgaggaaggaagag1283leuaspthrproproaspproalaleuaspgluvalarglysgluglu275280285tggccgctggtggacgactgcacgggagtcaccggctaccatgagcag1331trpproleuvalaspaspcysthrglyvalthrglytyrhisglugln290295300cttaccatccacggcaaggaccacgagagtgtgttcaccgtgtccctg1379leuthrilehisglylysasphisgluservalphethrvalserleu305310315tgggactgcgaccgcaagttcagggtcaagatcagaggcattgatatc1427trpaspcysasparglyspheargvallysileargglyileaspile320325330335cccgtcctgcctcggaacaccgacctcacagtttttgtagaggcaaac1475provalleuproargasnthraspleuthrvalphevalglualaasn340345350atccagcatgggcaacaagtcctttgccaaaggagaaccagccccaaa1523ileglnhisglyglnglnvalleucysglnargargthrserprolys355360365cccttcacagaggaggtgctgtggaatgtgtggcttgagttcagtatc1571prophethrglugluvalleutrpasnvaltrpleuglupheserile370375380aaaatcaaagacttgcccaaaggggctctactgaacctccagatctac1619lysilelysaspleuprolysglyalaleuleuasnleuglniletyr385390395tgcggtaaagctccagcactgtccagcaaggcctctgcagagtccccc1667cysglylysalaproalaleuserserlysalaseralagluserpro400405410415agttctgagtccaagggcaaagttcggcttctctattatgtgaacctg1715sersergluserlysglylysvalargleuleutyrtyrvalasnleu420425430ctgctgatagaccaccgtttcctcctgcgccgtggagaatacgtcctc1763leuleuileasphisargpheleuleuargargglyglutyrvalleu435440445cacatgtggcagatatctgggaagggagaagaccaaggaagcttcaat1811hismettrpglnileserglylysglygluaspglnglyserpheasn450455460gctgacaaactcacgtctgcaactaacccagacaaggagaactcaatg1859alaasplysleuthrseralathrasnproasplysgluasnsermet465470475tccatctccattcttctggacaattactgccacccgatagccctgcct1907serileserileleuleuaspasntyrcyshisproilealaleupro480485490495aagcatcagcccacccctgacccggaaggggaccgggttcgagcagaa1955lyshisglnprothrproaspprogluglyaspargvalargalaglu500505510atgcccaaccagcttcgcaagcaattggaggcgatcatagccactgat2003metproasnglnleuarglysglnleuglualaileilealathrasp515520525ccacttaaccctctcacagcagaggacaaagaattgctctggcatttt2051proleuasnproleuthralagluasplysgluleuleutrphisphe530535540agatacgaaagccttaagcacccaaaagcatatcctaagctatttagt2099argtyrgluserleulyshisprolysalatyrprolysleupheser545550555tcagtgaaatggggacagcaagaaattgtggccaaaacataccaattg2147servallystrpglyglnglngluilevalalalysthrtyrglnleu560565570575ttggccagaagggaagtctgggatcaaagtgctttggatgttgggtta2195leualaargarggluvaltrpaspglnseralaleuaspvalglyleu580585590acaatgcagctcctggactgcaacttctcagatgaaaatgtaagagcc2243thrmetglnleuleuaspcysasnpheseraspgluasnvalargala595600605attgcagttcagaaactggagagcttggaggacgatgatgttctgcat2291ilealavalglnlysleugluserleugluaspaspaspvalleuhis610615620taccttctacaattggtccaggctgtgaaatttgaaccataccatgat2339tyrleuleuglnleuvalglnalavallysphegluprotyrhisasp625630635agcgcccttgccagatttctgctgaagcgtggtttaagaaacaaaaga2387seralaleualaargpheleuleulysargglyleuargasnlysarg640645650655attggtcactttttgttttggttcttgagaagtgagatagcccagtcc2435ileglyhispheleuphetrppheleuargsergluilealaglnser660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( 2 ) information for seq id no : 2 :( i ) sequence characteristics :( a ) length : 1049 amino acids ( b ) type : amino acid ( d ) topology : linear ( ii ) molecule type : protein ( xi ) sequence description : seq id no : 2 : metgluleuileproilegluphevalleuprothrserglnarglys151015cyslysserprogluthralaleuleuhisvalalaglyhisglyasn202530valgluglnmetlysalaglnvaltrpleuargalaleugluthrser354045trpargglyleuleuproproalaglythralaserleuproproala505560leuserglugluglyglntrptyrgluiletyrasplystyrglnval65707580valglnthrleuaspcysleuargtyrtrplysalathrhisargser859095proglyglnilehisleuvalglnarghisproproserglugluser100105110glnalapheglnargglnleuthralaleuileglytyraspvalthr115120125aspvalserasnvalhisaspaspgluleugluphethrargarggly130135140leuvalthrproargmetalagluvalalaserargaspprolysleu145150155160tyralamethisprotrpvalthrserlysproleuproglutyrleu165170175trplyslysilealaasnasncysilepheilevalilehisargser180185190thrthrserglnthrilelysvalserproaspaspthrproglyala195200205ileleuglnserphephethrlysmetalalyslyslysserleumet210215220aspileprogluserglnsergluglnaspphevalleuargvalcys225230235240glyargaspglutyrleuvalglygluthrproilelysasnphegln245250255trpvalarghiscysleulysasnglyglugluilehisvalvalleu260265270aspthrproproaspproalaleuaspgluvalarglysgluglutrp275280285proleuvalaspaspcysthrglyvalthrglytyrhisgluglnleu290295300thrilehisglylysasphisgluservalphethrvalserleutrp305310315320aspcysasparglyspheargvallysileargglyileaspilepro325330335valleuproargasnthraspleuthrvalphevalglualaasnile340345350glnhisglyglnglnvalleucysglnargargthrserprolyspro355360365phethrglugluvalleutrpasnvaltrpleuglupheserilelys370375380ilelysaspleuprolysglyalaleuleuasnleuglniletyrcys385390395400glylysalaproalaleuserserlysalaseralagluserproser405410415sergluserlysglylysvalargleuleutyrtyrvalasnleuleu420425430leuileasphisargpheleuleuargargglyglutyrvalleuhis435440445mettrpglnileserglylysglygluaspglnglyserpheasnala450455460asplysleuthrseralathrasnproasplysgluasnsermetser465470475480ileserileleuleuaspasntyrcyshisproilealaleuprolys485490495hisglnprothrproaspprogluglyaspargvalargalaglumet500505510proasnglnleuarglysglnleuglualaileilealathrasppro515520525leuasnproleuthralagluasplysgluleuleutrphisphearg530535540tyrgluserleulyshisprolysalatyrprolysleupheserser545550555560vallystrpglyglnglngluilevalalalysthrtyrglnleuleu565570575alaargarggluvaltrpaspglnseralaleuaspvalglyleuthr580585590metglnleuleuaspcysasnpheseraspgluasnvalargalaile595600605alavalglnlysleugluserleugluaspaspaspvalleuhistyr610615620leuleuglnleuvalglnalavallysphegluprotyrhisaspser625630635640alaleualaargpheleuleulysargglyleuargasnlysargile645650655glyhispheleuphetrppheleuargsergluilealaglnserarg660665670histyrglnglnargphealavalileleuglualatyrleuarggly675680685cysglythralametleuhisaspphethrglnglnvalglnvalile690695700glumetleuglnlysvalthrleuaspilelysserleuseralaglu705710715720lystyraspvalserserglnvalileserglnleulysglnlysleu725730735gluasnleuglnasnserglnleuprogluserpheargvalprotyr740745750aspproglyleulysalaglyalaleualaileglulyscyslysval755760765metalaserlyslyslysproleutrpleugluphelyscysalaasp770775780prothralaleuserasngluthrileglyileilephelyshisgly785790795800aspaspleuargglnaspmetleuileleuglnileleuargilemet805810815gluseriletrpgluthrgluserleuaspleucysleuleuprotyr820825830glycysileserthrglyasplysileglymetilegluilevallys835840845aspalathrthrilealalysileglnglnserthrvalglyasnthr850855860glyalaphelysaspgluvalleuasnhistrpleulysglulysser865870875880prothrgluglulyspheglnalaalavalgluargphevaltyrser885890895cysalaglytyrcysvalalathrphevalleuglyileglyasparg900905910hisasnaspasnilemetilethrgluthrglyasnleuphehisile915920925asppheglyhisileleuglyasntyrlysserpheleuglyileasn930935940lysgluargvalprophevalleuthrproasppheleuphevalmet945950955960glythrserglylyslysthrserprohispheglnlyspheglnasp965970975ilecysvallysalatyrleualaleuarghishisthrasnleuleu980985990ileileleuphesermetmetleumetthrglymetproglnleuthr99510001005serlysgluaspileglutyrileargaspalaleuthrvalglylys101010151020asnglugluaspalalyslystyrpheleuaspglnilegluvaltrp1025103010351040glnargglnargmetaspcysalaval1045 ( 2 ) information for seq id no : 3 :( i ) sequence characteristics :( a ) length : 4137 base pairs ( b ) type : nucleic acid ( c ) strandedness : both ( d ) topology : linear ( ii ) molecule type : cdna ( vi ) original source :( a ) organism : homo sapiens ( ix ) feature :( a ) name / key : cds ( b ) location : 423 .. 3572 ( xi ) sequence description : seq id no : 3 : 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alaargpheleuleulysargglyleuargasnlys169016951700agaattggtcactttttgttttggttcttgagaagtgagatagcccag2435argileglyhispheleuphetrppheleuargsergluilealagln1705171017151720tccagacactatcagcagaggttcgctgtgattctggaagcctatctg2483serarghistyrglnglnargphealavalileleuglualatyrleu172517301735aggggctgtggcacagccatgctgcacgactttacccaacaagtccaa2531argglycysglythralametleuhisaspphethrglnglnvalgln174017451750gtaatcgagatgttacaaaaagtcacccttgatattaaatcgctctct2579valileglumetleuglnlysvalthrleuaspilelysserleuser175517601765gctgaaaagtatgacgtcagttcccaagttatttcacaacttaaacaa2627alaglulystyraspvalserserglnvalileserglnleulysgln177017751780aagcttgaaaacctgcagaattctcaactccccgaaagctttagagtt2675lysleugluasnleuglnasnserglnleuprogluserpheargval1785179017951800ccatatgatcctggactgaaagcaggagcgctggcaattgaaaaatgt2723protyraspproglyleulysalaglyalaleualaileglulyscys180518101815aaagtaatggcctccaagaaaaaaccactatggcttgagtttaaatgt2771lysvalmetalaserlyslyslysproleutrpleugluphelyscys182018251830gccgatcctacagccctatcaaatgaaacaattggaattatctttaaa2819alaaspprothralaleuserasngluthrileglyileilephelys183518401845catggtgatgatctgcgccaagacatgcttattttacagattctacga2867hisglyaspaspleuargglnaspmetleuileleuglnileleuarg185018551860atcatggagtctatttgggagactgaatctttggatctatgcctcctg2915ilemetgluseriletrpgluthrgluserleuaspleucysleuleu1865187018751880ccatatggttgcatttcaactggtgacaaaataggaatgatcgagatt2963protyrglycysileserthrglyasplysileglymetilegluile188518901895gtgaaagacgccacgacaattgccaaaattcagcaaagcacagtgggc3011vallysaspalathrthrilealalysileglnglnserthrvalgly190019051910aacacgggagcatttaaagatgaagtcctgaatcactggctcaaagaa3059asnthrglyalaphelysaspgluvalleuasnhistrpleulysglu191519201925aaatcccctactgaagaaaagtttcaggcagcagtggagagatttgtt3107lysserprothrgluglulyspheglnalaalavalgluargpheval193019351940tattcctgtgcaggctactgtgtggcaacctttgttcttggaataggc3155tyrsercysalaglytyrcysvalalathrphevalleuglyilegly1945195019551960gacagacacaatgacaatattatgatcaccgagacaggaaacctattt3203asparghisasnaspasnilemetilethrgluthrglyasnleuphe196519701975catattgacttcgggcacattcttgggaattacaaaagtttcctgggc3251hisileasppheglyhisileleuglyasntyrlysserpheleugly198019851990attaataaagagagagtgccatttgtgctaacccctgacttcctcttt3299ileasnlysgluargvalprophevalleuthrproasppheleuphe199520002005gtgatgggaacttctggaaagaagacaagcccacacttccagaaattt3347valmetglythrserglylyslysthrserprohispheglnlysphe201020152020caggacatctgtgttaaggcttatctagcccttcgtcatcacacaaac3395glnaspilecysvallysalatyrleualaleuarghishisthrasn2025203020352040ctactgatcatcctgttctccatgatgctgatgacaggaatgccccag3443leuleuileileleuphesermetmetleumetthrglymetprogln204520502055ttaacaagcaaagaagacattgaatatatccgggatgccctcacagtg3491leuthrserlysgluaspileglutyrileargaspalaleuthrval206020652070gggaaaaatgaggaggatgctaaaaagtattttcttgatcagatcgaa3539glylysasnglugluaspalalyslystyrpheleuaspglnileglu207520802085gtttggcagagacaaaggatggactgtgcagtttaattggtttctacatcttg3592valtrpglnargglnargmetaspcysalaval20902095ttcttggcatcaaacaaggagagaaacattcagcctaatactttaggctagaatcaaaaa3652caagttagtgttctatggtttaaattagcatagcaatcatcgaacttggatttcaaatgc3712aatagacattgtgaaagctggcatttcagaagtatagctcttttcctacctgaactcttc3772cctggagaaaagatgttggcattgctgattgtttggttaagcaatgtccagtgctaggat3832tatttgcaggtttggttttttctcatttgtctgtggcattggagaatattctcggtttaa3892acagactaatgacttccttattgtccctgatattttgactatcttactattgagtgcttc3952tggaaattctttggaataattgatgacatctattttcatctgggtttagtctcaattttg4012gttatctttgtgttcctcaagctctttaaagaaaaagatgtaatcgttgtaacctttgtc4072tcattccttaaatgatgcttccaaacatctccttagtgtctgcaggtgttagtggtgtgc4132taaaa4137 ( 2 ) information for seq id no : 4 :( i ) sequence characteristics :( a ) length : 1050 amino acids ( b ) type : amino acid ( d ) topology : linear ( ii ) molecule type : protein ( xi ) sequence description : seq id no : 4 : 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( 2 ) information for seq id no : 5 :( i ) sequence characteristics :( a ) length : 15 amino acids ( b ) type : amino acid ( d ) topology : linear ( ii ) molecule type : protein ( xi ) sequence description : seq id no : 5 : asnserglnleuprogluserpheargvalprotyraspprogly151015 ( 2 ) information for seq id no : 6 :( i ) sequence characteristics :( a ) length : 7 amino acids ( b ) type : amino acid ( d ) topology : linear ( ii ) molecule type : protein ( xi ) sequence description : seq id no : 6 : lysasnglyaspaspleuarg15 ( 2 ) information for seq id no : 7 :( i ) sequence characteristics :( a ) length : 5 amino acids ( b ) type : amino acid ( d ) topology : linear ( ii ) molecule type : protein ( xi ) sequence description : seq id no : 7 : hisileaspphegly15__________________________________________________________________________