a publikus adatbázisból hiányzó fehérjék azonosítási lehetõségei

A publikus adatbázisból hiányzó fehérjék azonosítási lehetõségei>gi|39578743|emb|CAE57166.1| Hypothetical protein CBG25105 [Caenorhabditis briggsae]MTIASKERRCARLLTQSDATSYFEKYKRVRSTDMKVQQCQSVAFNCDGTKLVCGAFDKKVSIANVDGGRLRFSWVGSSHTSSVEQVACSEKQPNMFASASSDRNVCIWDARQSKPTHRITNKVGNFFVSWSPCDEYLIFLDKDNRVNTVDVRNYKVVN>gi|39578744|emb|CAE57164.1| Hypothetical protein CBG25098 [Caenorhabditis briggsae]MVENWDTSYAQVEEALLKSWPKNKESCAQLLIGYFDYYSRFDFRNFVVQCRREMTLSKMEKEWPRPICVEDPFDLNHNLSSGVTKKMFVFIMKVFINSRAVFMSEKPKMTDHNHAGFHLMYRKVLLDKCHQGSAPSDRQCHSCHRIGHFYESCPFRMQGKDNRRRYTSNSTNNSYRSTTGSIITGRRQRILMVLEIDWAIIKG>gi|39578745|emb|CAE57162.1| Hypothetical protein CBG25096 [Caenorhabditis briggsae]NVLHVIPMFPMASFIGYKAHFCYGDQLKKISESSAHILEESVDSLVPLPPTLDDVRHRSKQLKEEESLANF>gi|39578747|emb|CAE57160.1| Hypothetical protein CBG25094 [Caenorhabditis briggsae]MNGFRKLDHLNSASLTLKHLLNNSTVHQSPQSAIAEFVDNSYDANAKNCSIEVYDTPNNERIEILDDGDGMTRSEALNIVKFGFSNKVDNAIGRYGMGLKSGGLYIGRDILLLTKKDDEETAVFISHSFLRAENTDEKVYIPSPSWKYGEAHVPTIEDAERFDDECGIINQYMSVEGYESFEQLFDKIPGEHGTLIIISKLQRDPRGELEINITGDKWDIQDIGDNLPPHKLSLRKYLEILYLNPKMAITLRGKDVYPRNVVDNWMARYTVEYNGDMSTESLNKTIHEIQAQKAQ>gi|39578748|emb|CAE57159.1| Hypothetical protein CBG25093 [Caenorhabditis briggsae]PKNMIPMENKNLYPISDEDEIDVLNSDDEEMLPEGSFHHQPDEIYMVQEDGEGYYDDDDEDDPLQEYQYVEEDGVL>gi|39578749|emb|CAE57158.1| Hypothetical protein CBG25092 [Caenorhabditis briggsae]MSIINLLEPFQYNFRTQIFANPNKFIRAVVLYNKSQIFESFQSSGHDTFGGETAISRMFLCPPDSKTNLLTARRSFRNYHHLSSSLCSDCRFNQKRQRRLAKKEWEKRRISTECLMKFGKEKREQLRNQSILTKSRIEAADQSLFRANEMSSNGDSFGVFQNIEGVGDVENDDDDDGFF>gi|39578750|emb|CAE57157.1| Hypothetical protein CBG25091 [Caenorhabditis briggsae]PTPSSTITSVPTTVSTFTYEPPPASTFTAEPTPSSTITSVQTTVSTLTNEPPPASTFTAEPTSSSTITSEPPPSSTITSEPTPSLTITSEPPPSLTITSEPPTPSTITSEHSPASTITSEPPPSSTIISEPPPSSTITSEPHPSSTITPVPPPASTIASEPTPSSTITSVPTTVSTFTTEPPPSSTFTAEPTPSSTITSEPTPSLTITSEPPPSSTITSEPPPSSTITSEPTPSSTIPSEPTPSLTITSEPPPSLTITSEPPPSLTITSEPPPSSTITSEPLPASTFTDEPTPSLPILSEPLPSSTITSDASPLQLSPLNHLHLQLSPLYLPQFQLSPLNLLRRQAFTAEPTPSLTITSEPPPSLTITSEPPPPSTITSEHSPASTITSETPPSSTITSEPPPSSTIISEPPPSSTITSEPTPSLTITSEPTPSLTITSEPPPSLTITSEPPPSSTITSEPLPASTITSVPTTVSTFTSEPPPASTFTAEPTPSSTITSELPPSSTITSEPTPSLTITSEPTPSLPITSEPSPSLTITSEPPPSLFCRHIALVRPLETR>gi|39578751|emb|CAE57156.1| Hypothetical protein CBG25090 [Caenorhabditis briggsae]MLENLLKENLLKENLLKENLLKENLLKEENLLKENLHEENLPKENLPKENLPKENFPKENLPKENLPKENLPKENLPKENFPKEYFPKENLHKENVRKENLPKENLPKENLPKEENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKENLPKKNLPKEENLPKENLPKENLPKENLPKENLPKENLPKEENLPKENLPKENFPKENLPKEENLPKEENLPKEENLPKEENLPKEENLPKENLPKENLPKKIFLKKKIFLKKKIFLKKIFLKKIFLKKIFLKKIFLKNLPKEENLPKEENLPKEENLPKEENLPKDHLPKDHL>gi|39578752|emb|CAE57154.1| Hypothetical protein CBG25088 [Caenorhabditis briggsae]KWIQWYLLRMLLQLKMGQRTVDILHQKGISIRILSKKNTSKIESIRYEYNVGYFDKQVQLMNWIIQMFRFSGEKIVVAIQRNSKERFLFRNRLRVTLWKRNAGGRRDYVPIHDASIPYPAHWPTDLIHFINAMLKFDKEKRLVGLEAIKKHAYTERIDFKSVFERKPAPVFIPCKEGLNCDPMYELEERILVSTPIHRRRTNHNNSSGRSSSEPQNAALVEVSKAFIDFSRHNMKIEPNGVCRSN>gi|39578754|emb|CAE57152.1| Hypothetical protein CBG25086 [Caenorhabditis briggsae]MEPKEPELTQEEIDHIAWIEKIAEQSSFEQMTAPPTRPPVPIKMATVDEPVVVTEAHEDEDRSSATSGADFQQSFDHEVTYERSSPLLEPDEVPVMEPKEPELTQEEIDHIAWIEKIAEQSSFEQMNAPPTRPPVPIKMATVDEPVVVTEAHEDEDRSSVTSGADFQQSFDHEVTYERSSPLLEPVEEPVMEPKEPELTQEEIDHIAWIQKIAEQSSFEQTTAPPTRPPVPIRMATVDEPNIVTEAHEDEDRSSATSGADFQQSFDHEVTYERSSPLLEPVEIPVKEPKEPELTQEEIDHIAWIEKIAEQSSFEQMTAPPTRPPVPIRMATVDEPNIVTEAHEDEHRSSATSGADFQQSFDHEVTYERSSPLLEPVEEPVMEPKEPQLPQEEIVHIAWIEKIAAQSSFEQMTAPPTRPPVPIRMAAVEEPNIVTEAHEDEDRSSATSGADFQQSFDHEVTYERSSPLLEPVEEPVMEPKEPELTQEEIDHIAWIEKIDEQSSFEQMTAPRTRPPVPIKMTTVDEPVVVTEAHEDEDRSSATSGADFQQSFDHEVTYERSSPLLEPVEIPVIEPKEPELTQEEIDHIAWIEKIAEQSSFEQMTAPPTRPPVPIRMAT>gi|39578755|emb|CAE57151.1| Hypothetical protein CBG25085 [Caenorhabditis briggsae]MTDGEESFKYYSKMKYLEDTQVLRCLVHRRNNLTFIQSKKNISQSDLNKIFGRQIGDEVENGIFNVMSSEAMEAEIVKYEKKEFCKSILTGKSELIRYANFKKTGILSAQSQVKALRKHGFRTFSLSHVFEMPKTVWRSVPKQMKIGEVFNKNVIYHKSGQYLVEKDDKSFFVVKTTETNIECKDPKCASKPAITCSHVFAVLSTLPTKKCQEVLNIREKFLISLSRSLQIKSINDGKDAKRYPNRKYSKRSHNNGGRTVSEFKKVNRNVSGESESGDDERKADGNISSDEENDSDCDVNSLNSKPNVFDESGDIVEDEISCNSNDEHVNLVLDPSVDHLNSHDSCMDLSDDPRKYDSDSDEENDIFGYQNVNPVPKPSNRVSNDHSSQIDLRDVLGEKGSRKPESDSEEVTDVSTGQHVKPMSLKEKFPIGYMHRLSDDSDDSET>gi|39578756|emb|CAE57150.1| Hypothetical protein CBG25084 [Caenorhabditis briggsae]MRFFFLLHILLMLHHFISHLLIQRAACGAHIADQVLEIMPFFDQHTELLSIHSVGPWRISFLISSVFSFFFFSAAFIFSTFLKVASPPEATADQKSRLDMLVQKVLNVGIDSKAQIQERTKEKVGKVMKETEEIKGKFMDIKKFTLADKRGKPIKEELEMEKKKRQMLLDEIKRLGEAKEEMSNDDDEPFENLMKEIDKLFEEADENRKLQKEADKEIATAATRQDAVFNEEMEEIEKFLSDIRNADKLQL>gi|39578757|emb|CAE57149.1| Hypothetical protein CBG25082 [Caenorhabditis briggsae]EFQNFRKTHKNSEFRNFRNLETQGSEKSEFQISGFRFQILYFRFQISDFRFQISDFRFQISDFRFQISDFRFQISDFRFQISDQISDFGEENVEASFYLSLKIFSEFQISGFQNFRKTHKNSEFRNFRNLETQGSEKFQISDFIFQISDFRFQISDFRFQISDFRFQISDFRFQISDFRFQISDFRFQISDFRFQISDQISDFGEENVEASFYLSLK

>gi|39578758|emb|CAE57148.1| Hypothetical protein CBG25081 [Caenorhabditis briggsae]MYSWTRSRYWLTRSMWRWIQSRYSWWTRSMYSWTRSRWRWIQSRYSWTRSRYSLTRSMWRWIQSRYLWTRSRYSWTRSMWRWIQSMYSWTRSRWRWIQSRYSWTRSRYSLTRSKWRWIQSRYSWTRSRYSWTRSMWRWIQSRYSWTRSRYSWTRSMWRWIQLRYSWTRSRYSLTRSRWRWIQSMYSWTRSRWRWIQSMYSWTQSRWRWIQSRYSWTRSRYSLTRSMWRWIQSRYSWTRSRYWLTRSMCRWIQSRYLWTRSRYSWTRSRWRWIQSRYSWTRSRWRWIQSRYSWTRSRYSWTRSKCRWIQSRYSWTRSRYSWTRSMWRWIQLRYSWTRSRYSWTRSMWRWIQSRYLWTRSRYSWTRSMWRWIQSMYSWTRSRWRWIQSRYSWTRSRYSLTRSKWRWIQSRYSWTRS>gi|39578759|emb|CAE56373.1| Hypothetical protein CBG24048 [Caenorhabditis briggsae]GLTEYVTVMACIVVDAEPIFGAIYRPFFNETIFGLQGFGVVTSDGKKISPVDEGKTDKKVVVSRSHAGKVKEIVEKVYGDKMTIEAAGGSGYKTLRLVNGTAGKGNKYIF>gi|39578760|emb|CAE57147.1| Hypothetical protein CBG25080 [Caenorhabditis briggsae]TKPNPQNMLIGKQKKAKAEGYDTNSLAIHSKVNASDFIKSSGYLDILGEANVILLDDPKWKNHEKTTEEVVEYMKDVKVLGPRELRVLLRWRKSMLETLEAERKAVEGEAQDVEIEEDLTEEQIEDRAMAEIDELIAKASEDEKAALKKKKKKMLKAKARVLKRRELKMIIDGDEGPQAEDQEVFQLKKIRKAKELAEITKETQAPDFENVEGKGAGWRGWGGREWLYLEISLFSAVF>gi|39578761|emb|CAE57145.1| Hypothetical protein CBG25077 [Caenorhabditis briggsae]MWAEPHHRISELFQNFRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISEFQNLRISESQNLRISEFQNLRISESQNLRISESQNLRISESQNLRISESQNLRISESQNLRISEFQNF>gi|39578762|emb|CAE56137.1| Hypothetical protein CBG23749 [Caenorhabditis briggsae]MPNRERYVKKFTVEQKRKMDEVFVASRNPEKGKIAALVAELELTTTQVRQYFRKQRFKNPIPEEIPRFLSEEQVEHGIVDGETFKRERNSEKRNQFEDAMEKFFEERQFLEEPDETLELESGGSRKRISDWLDKKRYDTLKSFLEKEISVLPNEMATFEKLSEKYCLDTTTHPDVILYIEMKED>gi|39578763|emb|CAE57144.1| Hypothetical protein CBG25076 [Caenorhabditis briggsae]GNFHNYVCSVNGTSPDPRYKRFINFSKKLRNCGIQVTMAQMFMFQKDENSFLDYNKERSPYFKFSEICITDWEKFWKKLLQFSCSLAGYSSSHLAFYSHDLARKVLKI>gi|39578764|emb|CAE57143.1| Hypothetical protein CBG25074 [Caenorhabditis briggsae]NGGYSHRSTLSRDLPSEETITSITPFDEEKLSDDSIDQSAQEISHSSKTDDMSDEKMSGVSISPIDPDESIQLTLPNEQEASRSSITEGIAEEMLPVDPIGTVAQGESVPSFPPTSFSDTIAQLRLYFNQELSNEGAMPAVSADRAIHSAKRTSSVPIRQYSSDTVDENELAVKKPRFFSPQRQVLKPLNTSYKLAMSPEAPSSEHPNSLVEPLVVLQKSPEEPWAELLNRPKTQTLPEALQQLRPLKRYGKQVTLKEYLQIQFKGQLLRTIDWNRVDDLKLFHPKTALFQSTNHPILLLAGKSFNRALLVEGNHRTAALSECHAQKELSDDDLKSTVPLTFFQIPKPEFNNMWMAANELGRQVQAQARFDWKNASSRTREAVFSEFKKEAYRTKGKFSKFESSIL>gi|39578765|emb|CAE57142.1| Hypothetical protein CBG25073 [Caenorhabditis briggsae]MVLYQMTLFFSVQMLFAIFLEYMPKTYCTDDDYCYKMKNKCLTDFIREDDSQICPYNSTDFKQCVLDAKRVDFRSAQFDYQQDCTGLKHFSSSTATFIGTLLGNLVLIYLSDTIGRRPVFIFSIALGVPALILSAAINGVMNFYIFRFIVGFAVAGTLTVGWTYSSEMITPSRRFRLRTFPNWA>gi|39578767|emb|CAE57140.1| Hypothetical protein CBG25071 [Caenorhabditis briggsae]MAFIFSTGYQMAFMQYGMTTIITLNRLTVLLNFTFFEPIWKKYSWLLVIFLIFLPFLSTKVALNYDSKFSYQNSTDYFLLTSGMPVWEIYSILIPFMIVTIVTSLAANLSSFVFVKHRKLVMAYKA>gi|39578768|emb|CAE57141.1| Hypothetical protein CBG25072 [Caenorhabditis briggsae]ITSITLIIQVIGNFLSIGMLTFEHSDYLPYLSIIRPFVSDGLTLVQPWLLYFFSSPMRKKLQQMFKRSSPATVATVVFKSRSV>gi|39578769|emb|CAE57139.1| Hypothetical protein CBG25067 [Caenorhabditis briggsae]MYGLGFVVNSAKLANARSLWFNVGALNHLHRTWSLGQGVVFSRAVLLSCDLLRLSFD>gi|39578770|emb|CAE56385.1| Hypothetical protein CBG24068 [Caenorhabditis briggsae]MKKITESGIRNPEISGFKNGSYPRTQVIATMVIPGNMPSAFPFFLTFKLVSSCSTSSFFHKSLGNLNFRSKSVTMAKGNIFVRFKNYIVETISDTYHHIRELRAEWNRQQGRG>gi|39578771|emb|CAE57138.1| Hypothetical protein CBG25066 [Caenorhabditis briggsae]SPFKDEAVFRFSNWFKKLGDEDKDKEAEEKEKELAEIAETRFLENKQKFRSLLIERKFSAEEWTLMKQFLDYPELAAIVVKMRRSQGVPGLRQNRRRCLTCEESLAGMPLTAHKTENCSIPYESRILVSKRLFGQRKCYGCGRMKEDGNCNCKSVPCRRCWRWNVPESFPHLQVMGTCQMVWKIRQETGHVPTEHEISDYTKRNDKIASIIQHKLEKFVAETERELVDAGRASNQKTEAELWWVADKYRTLAFSIPWHGAKGYFRKISKLFESPETI>gi|39578772|emb|CAE56541.1| Hypothetical protein CBG24269 [Caenorhabditis briggsae]MIEAVDRILVVDASIVTDTSIQKENLISSSLNQHGEINFVIETCDQYLHLHKSYIYLEVDLSKLDDTALAETDEVSLTNNAPMFLFDRATYSMDGVQVENIWILVELV>gi|39578773|emb|CAE57137.1| Hypothetical protein CBG25062 [Caenorhabditis briggsae]VLQTDESGNYIYPAVGPDGQVLPTDATGKTVYPVRGPDGTPLPTDASGAVIGPDGEPIPTDASGKPLSQDGSPLPTDNQGNYVLVPTGEDVTKYLPTDESGNVIYPITRPDGTPLGTDASGNFVTDDGTVVEKDDEGKPVGPDGAVLPTDASGNYIYPVVGPDGQVLPTDATGKTVYPVRGPDGTPLPTDGSGAVIGPDGEPIPTDPSGKPLSKDGSPLPTDASGNYILVPAEEETTKTLPTDSEGNVIYPITNPDGTPLGTDSSGAFVTQDGTVIEKNEDGKPVGPDGQFVYSVIADIFFPYLQNQGPVLQISKKN>gi|39578774|emb|CAE57136.1| Hypothetical protein CBG25061 [Caenorhabditis briggsae]MGMKWTGLTFEIAAEEMRRLKSTPSEREQMKLYGLYKQALHGDIPNEDVYPVSQKEKQSLEPEKSSALAERVYLTN>gi|39578775|emb|CAE56809.1| Hypothetical protein CBG24622 [Caenorhabditis briggsae]SSFFGMLVYCVRVCMYVNDTEKEGEREELSMSYNQKRYTPKRRRNEIDDMMNAGQMAVGSSGTIAMGTGVPQKKDDMDALSTLERDVAKRTQHAQQILLRQRANQSTPRSVTPPPQPQEFKTYVISAPQQQHQQHVGNNGHQQVLHGGHMQQQQQIQVSHQEIMSEKAKFGSANVARVSLRLHWPAKYLADHFFV

NO MATCH!

Csorba Attila esettanulmánya az adatbázisokkal

ZULU 20050818 1123

ZULU 20050818 1123

Célpont: Capsicum annuum kitinázaktivitást mutató fehérjéi,aktivitásfestés alapján, 1D SDS-PAGE gélcsíkok,(Ca1, Ca2, Ca3, Ca4, Ca5, 3220kDa)

Probléma: Capsicum annuum, alias paprikaA fehérjeadatbázis defektív az alanyra nézve.

Bizonyítás: Capsicum annuum vs. Oryza sativa(paprika kontra rizs)

Fehérjetalálatok száma az NCBInr 2006.04.20-i adatbázisban:

675 121910

Célpont: Capsicum annuum kitinázaktivitást mutató fehérjéi,aktivitásfestés alapján, 1D SDS-PAGE gélcsíkok,(Ca1, Ca2, Ca3, Ca4, Ca5, 3220kDa)

Probléma: Capsicum annuum, alias paprikaA fehérjeadatbázis defektív az alanyra nézve.

Bizonyítás: Capsicum annuum vs. Oryza sativa(paprika kontra rizs)

Fehérjetalálatok száma az NCBInr 2006.04.20-i adatbázisban:

675 121910>3.5 millió (6×10-18mol)

Felszerelés:#Bruker Reflex III, MALDI-TOF

nagy felbontás (>10000), pontosság (∆m<200ppm),szimpla minták jó azonosíthatósága (PSD)

#Agilent 1100 series, LC/MSD Trap, HPLC-oszlop: Zorbax 300SB-C18, 3.5µm, 150mm×75µm komplex minták könnyű azonosítása, nagyszámú MS/MS adat

Stratégia:MALDI-TOF:PMF

Nincs/kevés fehérje PMF-lekeresés

A spektrum minősége

Eredményesség

PSD alátámasztás Komplex minta és/vagydefektív adatbázis

Sikeresség

Gotcha!Komplex minta és/vagy

defektív adatbázis

LC-MS/MS?

B-terv: LC-MS/MS

Nincs/kevés fehérje MS/MS lekeresés

MS/MS információ

Gotcha! Defektív adatbázis

nonredundáns PDB

Homológia keresés(BLAST)

Manuális szekvenálás

Saját adatbázis létrhozása

Mission possible?!

m/z1000 1200 1400 1600 1800 2000 2200 2400

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

[Abs. Int. * 1000]

MALDI-TOF, Ca1:

PDB lekeresés:1. gi|72534134 peroxidase POA1 [Capsicum annuum] 2. gi|17066703 peroxidase [Capsicum annuum]

m/z1000 1200 1400 1600 1800 2000 2200 2400

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

[Abs. Int. * 1000]

965.483242-249

968.55361-69

1073.581108-117

1089.58912-21

1173.5282-11

1288.625199-210

1624.84722-36

1883.903125-142

2355.39279-100

1.

m/z1000 1200 1400 1600 1800 2000 2200 2400

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

[Abs. Int. * 1000]

965.483276-283

968.55395-103

1073.581142-151

1089.58946-55

1173.52836-45

1288.625233-244

1883.903159-176

2220.228245-264

2355.392113-134

2.

Mi a különbséga szekvenciák

között?

Akkor mostmelyik?

Válasz:

PSD analízis!

Ca1: Lefedettségi térkép, szekvenciák összehasonítása

RVGFYS STCPRAESIV

QSTVRSHFQS DPTVAPGLLR MHFHDCFVQG CDASILISGS GTERTAPPNS

LLRGYEVIDD AKQQIEAICP GVVSCADILA LAARDSVVVT RGLTWSVPTG

RRDGLVSRAS DTSDLPGFTE SVDSQKQKFS AKGLNTQDLV TLVGGHTIGT

SACQFFSYRL YNFNSTGGPD PSIDASFLPT LRGLCPQNGD GSKRVALDTG

SVNNFGTSYF SNLRNGRGIL ESDQKLWTDD STKVFIQRYL GLRGFLGLRF

GVEFGRSMVK MSNIEVKTGT NGEIRKVCS

1. gi|72534134

MEYYYNYNSI NKMVSIIFIL VLAIDLTMVL GQGTRVGFYS STCPRAESIV

QSTVRSHFQS DPTVAPGLLT MHFHDCFVQG CDASILISGS GTERTAPPNS

LLRGYEVIDD AKQQIEAICP GVVSCADILA LAARDSVLVT KGLTWSVPTG

RRDGLVSRAS DTSDLPGFTE SVDSQKQKFS AKGLNTQDLV TLVGGHTIGT

SACQFFSYRL YNFNSTGGPD PSIDASFLPT LRGLCPQNGD GSKRVALDTG

SVNNFDTSYF SNLRNGRGIL ESDQKLWTDD STKVFIQRYL GLRGFLGLRF

GVEFGRSMVK MSNIEVKTGT NGEIRKVCSA IN

2. gi|17066703

Ca1: PSD analízis, m/z: 1624,85

m/z1000 1200 1400 1600 1800 2000 2200 2400

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

[Abs. Int. * 1000]

965.483242-249

968.55361-69

1073.581108-117

1089.58912-21

1173.5282-11

1288.625199-210

1624.84722-36

1883.903125-142

2355.39279-100

1.

m/z: 1624,85

m/z100 200 300 400 500 600 700 800 900

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

1.8

2.0

2.2

2.4

2.6

2.8

[Abs. Int. * 1000]

y 1

b 2

H

b 3

b 4

b 5

y 5

y-17 5

b 6

y 6

y-17 6

P

y 9

L

b-ionok \\\\\ SHFQSDPTVAPGLLR \\ \ y-ionok MALDI-TOF

alapján az erőshomológiaellenére:

1. gi|72534134peroxidase POA1[Capsicum annuum]

Ca2:

Ca5:Ca4:

Ca3:

m/z900 1000 1100 1200 1300 1400 1500 1600 1700 1800

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

[Abs. Int. * 1000]

1155.748147-157

956.474

995.523

997.603

1109.622

1165.574

1281.723

1358.731

1416.678

1430.719

1525.566

1588.836

1743.876

m/z1000 1200 1400 1600 1800 2000 2200 2400 2600 2800

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

[Abs. Int. * 1000]

970.627668-675

995.57330-38

1061.563739-748

1109.631309-320

1254.634667-676

1261.634516-527

1318.599736-747

1462.768503-515

1656.774337-351

1842.99639-55

1891.028691-708

2819.53630-55

907.548

919.517

955.518

967.556

1015.571

1042.590

1237.680

1251.585

1285.674

1302.617

1345.615

1358.711

1390.695

1426.667

1477.722

1588.823

1658.769

1743.8562206.253

m/z1000 1200 1400 1600 1800 2000 2200 2400 2600

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

[Abs. Int. * 1000]

1254.60321-34

1588.774610-629

1890.952579-601

2201.170381-399

955.476

967.594

995.550

1015.535

1034.651

1061.532

1072.537

1109.613

1157.569

1187.569

1210.567

1222.568

1229.595

1230.598

1251.590

1261.619

1281.681

1295.692

1302.561

1345.594

1358.685

1426.627

1427.631

1477.678

1574.784

1575.787

1627.686

1661.852

1664.731

2258.150

2518.390

m/z1000 1200 1400 1600 1800 2000 2200 2400 2600 2800

0

1

2

3

4

5

6

7

8

9

10

11

12

13

[Abs. Int. * 1000]

997.65367-74

1244.5731-10

1254.594164-174

1574.74480-93

1588.821172-184

1709.838207-220

1743.79043-57

2653.30194-118

991.613

995.540

1119.651

1157.620 1187.584

1210.599

1222.584

1313.682

1661.879

1903.065

Ca2:Species Protein Name

1 TOXOPLASMA GONDII casein kinase I beta isoform

2 BORDETELLA PARAPERTUSSIS LysR-family transcriptional regulator

3 LISTERIA MONOCYTOGENES STR. 4B F2365 oxidoreductase, Gfo/Idh/MocA family

4 LISTERIA MONOCYTOGENES STR. 4B H7858 oxidoreductase, Gfo/Idh/MocA family

5 AGROBACTERIUM TUMEFACIENS STR. C58 AGR_L_644p

6 BURKHOLDERIA PSEUDOMALLEI 1710A COG1562: Phytoene/squalene synthetase

7 BURKHOLDERIA PSEUDOMALLEI S13 COG1562: Phytoene/squalene synthetase

8 BURKHOLDERIA PSEUDOMALLEI 1655 COG1562: Phytoene/squalene synthetase

9 BURKHOLDERIA PSEUDOMALLEI 668 COG1562: Phytoene/squalene synthetase

10 BURKHOLDERIA MALLEI NCTC 10247 COG1562: Phytoene/squalene synthetase

11 BURKHOLDERIA PSEUDOMALLEI K96243 putative squalene/phytoene synthase

12 BURKHOLDERIA MALLEI ATCC 23344 squalene/phytoene synthase family protein

13 SOLIBACTER USITATUS ELLIN6076 von Willebrand factor, type A

14 BURKHOLDERIA PSEUDOMALLEI 1710B squalene/phytoene synthase family protein

15 CAENORHABDITIS BRIGGSAE Hypothetical protein CBG12555

16 CRYPTOSPORIDIUM PARVUM heat shock protein 60

17 CLOSTRIDIUM BEIJERINCKI NCIMB 8052 glucuronate isomerase

18 UNREADABLE gi|606946|gb|AAA64268.1| neural-restrictive silencer factor

19 MYCOPLASMA CAPRICOLUM SUBSP. CAPRICOLUM ATCC 27343 N utilization substance protein B (nusB), putative

20 PLASMODIUM BERGHEI STRAIN ANKA DNA-directed RNA polymerase II


1 MUS MUSCULUS adenylate kinase 3

2 PSEUDOMONAS SYRINGAE PV. PHASEOLICOLA 1448A exopolysaccharide biosythesis protein

3 CHLOROFLEXUS AURANTIACUS J-10-FL Acyl-CoA dehydrogenase, C-terminal:Acyl-CoAdehydrogenase, central region:Acyl-CoAdehydrogenase, central region

4 LEGIONELLA PNEUMOPHILA STR. LENS hypothetical protein

5 ERYTHROBACTER LITORALIS HTCC2594 ribosomal protein L10

6 ARABIDOPSIS THALIANA AT4g22240/T10I14_70

7 NOSTOC SP. PCC 7120 alr3497

8 LEISHMANIA MAJOR hypothetical protein, unknown function

9 PROCHLOROCOCCUS MARINUS STR. MIT 9211 rubredoxin

10 CAENORHABDITIS ELEGANS Hypothetical protein C52E2.5

11 CANIS FAMILIARIS PREDICTED: similar to C-type lectindomain family 14, member A

12 HERBASPIRILLUM SP. B501 nitrogenase Mo-Fe protein alpha chain

13 ORYZA SATIVA (JAPONICA CULTIVAR-GROUP) myb family protein-like

14 DANIO RERIO hypothetical protein LOC393878

15 LEGIONELLA PNEUMOPHILA STR. LENS hypothetical protein

16 DANIO RERIO T-cell receptor gamma

17 METHYLOBACILLUS FLAGELLATUS KT Protein of unknown function DUF1568

18 ASHBYA GOSSYPII ATCC 10895 ACL096Wp

19 ANOPHELES GAMBIAE STR. PEST ENSANGP00000011927

20 DEBARYOMYCES HANSENII CBS767 unnamed protein product


1 VARIOLA VIRUS A18R

2 VARIOLA MINOR VIRUS A19R protein

3 DANIO RERIO hypothetical protein LOC394103

4 DROSOPHILA MELANOGASTER CG33455-PA

5 DESULFITOBACTERIUM HAFNIENSE DCB-2 Helix-turn-helix, Fis-type

6 HAHELLA CHEJUENSIS KCTC 2396 ABC-type uncharacterized transport system, ATPase component

7 AZOARCUS SP. EBN1 putative membrane-bound regulator HflC

8 HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 envelope glycoprotein

9 NICOTIANA TABACUM NtPRp27

10 RATTUS NORVEGICUS PREDICTED: similar to BC021442 protein

11 XENOPUS LAEVIS MGC84728 protein

12 RHODOPSEUDOMONAS PALUSTRIS HAA2 transcriptional regulator, GntR family

13 SILICIBACTER POMEROYI DSS-3 choline sulfatase, putative

14 XANTHOMONAS AXONOPODIS PV. CITRI STR. 306

phosphoenolpyruvate carboxylase

15 MUS CAROLI eosinophil-associated ribonuclease 2 precursor

16 HELICOBACTER HEPATICUS ATCC 51449 glutamyl-tRNA synthetase

17 ORYZA SATIVA (JAPONICA CULTIVAR-GROUP) putative eukaryotic initiation factor (iso)4F subunit p82-34 (eIF-(iso)4F p82-34)

18 XENOPUS LAEVIS Xrcc1-prov protein

19 PLASMODIUM YOELII YOELII STR. 17XNL hypothetical protein PY02997

20 DANIO RERIO PREDICTED: hypothetical protein XP_696923

MALDI-TOF összefoglalás:

Ca1: PSD-vel igazolt paprikafehérjeCa2, Ca3, Ca4, Ca5: A lekeresések eredménye nem bíztató,

PSD analízisre alkalmatlan minták

•Ca2, Ca5: Gyenge spektrumok•Ca3, Ca4: Túl komplex minták

B-terv:

LC-MS/MS

LC-MS/MS eredmények az NCBInr adatbázisából:

Protein MW (Da) Species Protein Name

25264.6 Atropa belladonna putative NtPRp27-like protein

21400 Capsicum annuum leucine-rich repeat protein

27768.5 Capsicum annuum chitinase class II

31873 Capsicum annuum peroxidase POA1

36074 Capsicum annuum peroxidase

26583.7 Lycopersicon esculentum chitinase

34924.3 Lycopersicon esculentum peroxidase

34940.3 Lycopersicon esculentum peroxidase

68912.7 Lycopersicon esculentum LEXYL2

10663.9 Nicotiana tabacum putative basal resistance related chitinase

27378 Nicotiana tabacum NtPRp27

25151.5 Oryza sativa (indica cultivar-group) PR-3 class IV chitinase

30451.4Oryza sativa (japonica cultivar-group) OSJNBb0091E11.8

27040.1 Solanum chacoensetriose phosphate isomerase cytosolic isoform

25131.2 Solanum tuberosum NtPRp27-like protein

Egy kis növényrendszertan:

Eukaryota

Viridiplantae

Streptophyta

Streptophytina

Embryophyta

Tracheophyta

Euphyllophyta

Spermatophyta

Magnoliophyta

Eudicotyledons

Core Eudicotyledons

Asterids

Lamiids

Solanales

Solanaceae

Liliopsida

Commelinids

Poales

Poaceae

Ehrhartoideae

Oryzeae

Oryza

Oryzasativa

Solanum

Solanumtuberosum

Solanumchacoense

Lycopersiconesculentum

Capsicum

Capsicumannuum

Nicotiana

Nicotianatabacum

Atropa

Atropabelladonna

tBLASTn: TIGR adatbázis (genom adatbázis, 6 frame fordítás)http://www.tigr.org/

Species Protein Name

Capsicum annuum

TC3404 ORF64-827 frame +1 homologue to Q6T379 Triose phosphate isomerase cytosolic isoform- complete

Capsicum annuum

TC3587 ORF13-750 frame +1 homologue to CHIB_LYCES Q05540 cidic 27 kDa endochitinase precursor- part

Capsicum annuum TC3666 ORF97-506 frame +1 similar to Q7XB39 Class IV chitinase- partial

Capsicum annuum TC3667 ORF72-481 frame +3 similar to Q43151 Pathogenesis-related protein- PR-3

Capsicum annuum TC3709 ORF66-1063 frame +3 Q8W4V8 Peroxidase- complete

Capsicum annuum TC4206 ORF15-658 frame +3 homologue to O82552 Chitinase class II- partial

Capsicum annuum TC4719 ORF11-610 frame +2 homologue to Q84XQ4 NtPRp27-like protein- partial

Capsicum annuum TC5120 ORF29-789 frame +2 O82552 Chitinase class II- complete

Capsicum annuum

TC6065 ORF58-899 frame +1 similar to PERX_NICSY Q022 Lignin forming anionic peroxidase precursor – partial

Mission completed

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