Karl Clauser Proteomics and Biomarker Discovery Sample Experiment Summary 1 Spider 1 ug venom Reduce/alkylate De-salt LC-MS/MS 300ng C18 300A Microsorb

Karl ClauserProteomics and Biomarker Discovery 1

Sample Experiment SummarySpider

1 ug venom

Reduce/alkylate

De-salt

LC-MS/MS 300ngC18 300A Microsorb 3um

Orbitrap Elite CID/HCD/ETD

Spectrum Mill

Cysiodoacetamide

CysMeAziridine

DigestLys-C

Database• Venom gland mRNA library• Ion Torrent sequencing

• 75-150 bp reads• Transcriptome assembly (Trinity)• Translation (extractORFS.pl)

Database• Venom gland mRNA library• 454 sequencing

• 300-400 bp reads• Transcriptome assembly (MIRA)• Translation (extractORFS.pl)

Centipede 1 ug venom

Reduce/alkylateCys-ethanolyl-gp

Digest500ng & 500 ng Trypsin Glu-C

De-salt

LC-MS/MS 150 ng300A Microsorb 3um C18

C18 120A Reprosil 3umOrbitrap Elite CID/HCD/ETD

Spectrum Mill

Scorpion 1 ug venom

Reduce/alkylate

De-salt

LC-MS/MS 300,600 ngC18 300A Microsorb 3um

Orbitrap Elite CID/HCD/ETD

Spectrum Mill

Cys-ethanolyl-solCys-iodoacetamide

C18 200A Magic 3um

Cys-MeAziridine

DigestLys-C

Database• Venom gland mRNA library• 454 sequencing

• 300-400 bp reads• Transcriptome assembly (Trinity)• Translation (extractORFS.pl)


K121115_Hi_dsrak#Spectra

K121115_Hi_dsrakIntens i ty

K121115_Hi_dsrak#peps

K120223_Hi#Spectra

K120223_HiIntens i ty

K120223_Hi#peps

both#peps

scoreUnique subgroupNum entry_name BLAST2GO-Protein-Name

mature#Cys

cyspattern accession_number

SpiderPSignal peptide:

SpiderPPropeptide - MS/MS covered

80 2.75E+10 17 169 1.31E+10 38 53 775.68 2.1 unknown omega-hexatoxin-hi2a 6 37_1680.3d.comp485_c0_seq1_F3_R4 MKFSKLSLTLALILTQALFVLC GKINGGELLLHFPDFMENGLESHALHDEIRKPIDTEKADAER

61 4.03E+09 16 141 7.25E+09 35 49 706.92 2.2 omega-hexatoxin-Hi2b omega-hexatoxin-hi2b 6 AS58251 2.02E+09 15 124 3.90E+09 29 42 588.62 2.3 omega-hexatoxin-Hv2i omega-hexatoxin-hi2b 6 AS126353 2.06E+09 15 120 4.23E+09 26 39 553.17 2.4 omega-hexatoxin-Hv2n omega-hexatoxin-hi2b 6 AS127146 1.59E+09 13 120 4.38E+09 26 37 540.07 2.5 omega-hexatoxin-Hv2h_1 omega-hexatoxin-hi2b 6 AS126229 1.62E+09 9 112 2.56E+09 25 33 459.03 2.6 omega-hexatoxin-Hv2l omega-hexatoxin-hi2a 6 AS126938 1.41E+09 10 100 1.73E+09 21 29 435.72 2.7 omega-hexatoxin-Hv2o omega-hexatoxin-hi2a 6 AS1272

3 1.08E+07 1 66 2.03E+09 9 9 129.68 2.8 omega-hexatoxin-Hv2f omega-hexatoxin-hi2b 6 AS125812 1.02E+09 3 107 9.33E+09 25 28 431.57 6.1 unknown hntx- precursor 6 37_1680.3d.comp492_c0_seq1_F1_R2 MNTLIAFAVLLLLSTTLG DTDDKVSHEEIQERKELSGISEELLLQQLEAVEAALMEKERLEEMEEDGNSREKR

3 2.69E+06 1 82 4.05E+10 19 20 266.31 9.1 omega-hexatoxin-Hi1a_2 omega-hexatoxin-ar1a 6 AS192.310 2.01E+08 2 54 1.35E+10 13 15 204.53 9.2 omega-hexatoxin-Hi1g_1 omega-hexatoxin-ar1a 6 AS747

6 1.00E+08 2 59 2.06E+10 13 15 202.12 9.3 unknown omega-hexatoxin-ar1a 6 37_1680.3d.comp499_c1_seq1_F4_R1 MNTATGFIVLLVLATVIGCISA DFQGAFEPHEEEEAERIFRR

3 2.69E+06 1 48 7.22E+09 11 12 147.39 9.4 unknown omega-hexatoxin-ar1a 6 spider.q17.comp1689_c0_seq1_F1_R1 MNTATGFIVLLVLATVIGCISA DFQGAFEPHEEEEAERIFRR

3 2.69E+06 1 50 1.47E+10 10 11 150.20 9.5 omega-hexatoxin-Hi1b_8 omega-hexatoxin-ar1d 6 AS195.43 2.69E+06 1 40 1.20E+08 9 10 119.99 9.6 omega-hexatoxin-Hi1c_2 omega-hexatoxin-ar1e 6 AS197.2

26 2.40E+08 6 6 80.01 17.1 unknown mu-hexatoxin-mg1a 6 spider.q17.comp522_c0_seq2_F5_R2 MRVTLVFLVLSALAVTHG DTNLDMERKEAEESSLSEMKEMLLLQELQAVEAALFGKMNAENDENRDFREKR

58 7.92E+09 2 2 47.56 18.1 unknown 6 37_1680.3d.comp125_c0_seq1_F2_R1 MDVKTVLIFTVLFGVISNNAA None predicted

5 8.62E+07 2 2 31.60 22.1 unknown 6 37_1680.3d.comp466_c0_seq1_F4_R3 MNTMIGFIVLLVSATVLG DPELDALRKELDERFDMEFKDGPMSEIQEKLFLQELEALDSELLEDDPQTDTYENSNFREKR

2 5.66E+06 1 1 20.87 25.1 unknown 6 spider.q17.comp522_c0_seq1_F5_R2 MRVTLVFLVLSALAVTHG DTNLDMERKEAEESSLSEMKEMLLLQELQAVEAALFGKMNAENDENRDFREKR

6 3.77E+07 1 1 18.55 27.1 unknown 6 37_1680.3d.comp183_c0_seq1_F1_R19 1.27E+08 2 2 17.69 28.1 unknown 6 37_1680.3d.comp496_c0_seq1_F6_R2 MKITIFCLTILTLSITFG ETNEDVKDGSLKEMDEMSLLQELMSVEAALVEKEMEAEAEENRDSREKR

117 8.55E+09 20 381 4.84E+10 87 107 1647.02 1.1 unknown hntx-xviii precursor 8 CECCG 37_1680.3d.comp348_c0_seq1_F5_R3 MKLLYFFVVITVLVAVAAA LPAKTEEQIAAEENQLVEDLVQYAGTRLTQKRATR

30 1.22E+09 5 17 8.68E+08 7 12 174.31 11.1 unknown hntx-xviii precursor 8 CECCG 37_1680.3d.comp159_c0_seq1_F1_R13 8.62E+07 1 1 15.22 29.1 unknown 8 CECCG 37_1680.3d.comp5507_c0_seq1_F5_R1

32 4.96E+09 12 12 173.17 12.1 unknown hntx- precursor 8 CECCG spider.q17.comp2075_c0_seq1_F2_R2 MKLYLVILVTSVALAAA SPTRTKEEPIEDELLEALLSVEKSLFNEETTVMEKR

79 3.20E+09 14 22 1.53E+09 7 21 315.67 8.1 unknown u1-cyrtautoxin-as1a 8 CC,CYIC 37_1680.3d.comp410_c0_seq1_F2_R3 MKTILSMLIFVALFAAIVG None predicted

4 4.10E+07 2 2 25.46 24.1 unknown 8 CC,CYIC 37_1680.3d.comp429_c0_seq1_F3_R3 MKLILLIAIFSALAVVNLG None predicted

13 7.13E+08 5 41 4.80E+09 9 14 163.06 13.1 unknown 8 CC,CXC,CXC 37_1680.3d.comp454_c0_seq1_F4_R5 MRIHYLLILSFLLLASG VLINSEEMPRSEMSPSR

16 5.68E+08 4 7 3.99E+08 3 7 109.57 15.1 unknown 8 CC,CXC,CXC 37_1680.3d.comp454_c0_seq2_F4_R5 MRIHYLLILSFLLLASG VLINSEEMPRSEMSPSR

3 5.71E+07 2 3 7.23E+07 1 3 40.15 21.1 unknown 8 CC,CXC,CXC spider.q17.comp1562_c0_seq3_F5_R4 MRIYSLLILSFLLLASTVLINS AEMPRSEKSLLYSIMQGREDSEEGR

7 1.56E+08 2 2 19.15 26.1 unknown 8 CC,CXC,CXC spider.q17.comp479_c0_seq2_F6_R2 MRISTLIVLSGLLLTTSVLIRA EEIPGLEESLVPAGLVDEESSPEQGR

37 1.07E+10 9 78 1.45E+10 25 32 541.87 5.1 unknown u2-hexatoxin-hi1a 8 CC,CXC,CXC 37_1680.3d.comp516_c0_seq1_F2_R2 MRNTTFLVLNVMLLVSVALFCAA DPEMEKSSFAEILDTGNPEQER

6 1.39E+08 4 4 41.24 20.1 unknown 8 CC,CXC,CXC spider.q17.comp1557_c0_seq1_F3_R1 MRNTTFLVLSALVLVSVALLCES APQMSEVDLVQSLEENPEGER

41 6.11E+09 8 51 1.41E+09 18 26 340.73 7.1 unknown delta-hexatoxin-mg1b 8 CCC 37_1680.3d.comp616_c0_seq1_F1_R1 None predicted MEAIEDAFLEKGFLPHEEDRNARPKR

3 2.75E+07 1 20 1.11E+09 5 6 116.98 14.1 unknown delta-hexatoxin-mg1b 8 CCC spider.q17.comp6008_c0_seq1_F1_R15 2.31E+08 3 16 1.30E+08 5 8 80.81 16.1 unknown delta-hexatoxin-mg1b 8 CCC spider.q17.comp1543_c0_seq2_F2_R1 MKIIIALYVLFFLTIALG DITEGNEDDLVDNFRKELSEADIPLLKKLEAIEDALLEKDFLPYEEEDRNARPKR

4 4.55E+07 2 2 13.28 30.1 unknown delta-hexatoxin-mg1b 8 CCC spider.q17.comp1516_c0_seq1_F6_R1168 3.01E+10 21 79 1.58E+10 21 39 643.52 3.1 unknown u1-hexatoxin-hsp201a 10 spider.q17.comp1699_c0_seq2_F3_R3 MLKFVVVIFLVIMASTFA None predicted

41 2.47E+09 10 25 6.24E+08 9 17 274.67 3.2 unknown u1-hexatoxin-hsp201c 10 spider.q17.comp1699_c0_seq3_F3_R3 MLKFVVVIFLVIMASTFA None predicted

66 2.06E+10 14 63 1.26E+10 20 34 579.80 4.1 unknown u1-hexatoxin-hi1a 10 37_1680.3d.comp452_c0_seq2_F2_R3 MLKFVVVIFLVIMASTFA None predicted

22 3.27E+08 8 27 7.84E+09 10 16 227.62 10.1 unknown hntx-xiv- precursor 10 37_1680.3d.comp799_c0_seq1_F2_R4 MMKAVIILVILFNAVYVSV None predicted

6 3.37E+08 2 2 2.00E+08 1 3 45.22 19.1 P81803 U1-hexatoxin-Hv1a u1-hexatoxin-hv1a 10 AS2093 9.42E+07 2 2 9.86E+07 1 3 29.83 23.1 unknown ?6,5 CC,CC spider.q17.comp1681_c0_seq3_F5_R6 MKLRITLALTSVLAFCVFG DKENENLMENLLEDDLLDIFTDAIHMER

Spider Protein Group.subgroup ListMatches to Arachnoserver but not RNA-SeqSuggests need to improve paralog inclusion inRNA-Seq Transcript Assembly

Samples are fromDifferent animals

Nov 2012 Feb 2012MeAzCys IAAcysLys-C undigested


Group 2 omega-hexatoxin Hi2

6 Cys

SubGroup

11215346666

MTPSCTLGICAPSVGGLVGGLLG reads: 4382MTPSCTLGICAPSVGGIVGGLLG reads: 0251MTPSCTLGICAPNVGGLVGGLLG reads: 0201MTPSCTMGLCVPNVGGLVGGILG reads: 0000MTPSCTMGICVPNVGGLVGGILG reads: 0000MTPSCTMGICVPNVGGLVGGLLG reads: 0000 CAPNVGG reads: 0442 CVPNVGG reads: 0000

Evidence exists in spider.q17 reads for paralogs missing from the assembly.

ERGVLDCVVNTLGC reads: 635ERGVVDCVLNTLGC reads: 106ERGLVDCVLNTLGC reads: 553ERGLADCVLNTLGC reads: 21ER_QLDCVLNTLGC reads: 26ERGVVGCVLNTLGC reads: 8

VLDCVVNTLGCSSDKDCCGMTPSCTLGICAPSVGGL reads: 99VVDCVLNTLGCSSDKDCCGMTPSCTLGICAPSVGGL reads: 33VVDCVLNTLGCSSDKDCCGMTPSCTLGICAPSVGGL reads: 33LVDCVLNTLGCSSDKDCCGMTPSCTLGICAPSVGGL reads: 72LADCVLNTLGCSSDKDCCGMTPSCTLGICAPSVGGL reads: 4VLDCVVNTLGCSSDKDCCGMTPSCTLGICAPNVGGL reads: 2QLDCVLNTLGCSSDKDCCGMTPSCTLGICAPSVGGL reads: 1QLDCVLNTLGCSSDKDCCGMTPSCTLGICAPNVGGL reads: 2


Only 1 spectrum supports LCVPN

(G)L C V P n\V\G\G\L\V\G\G\I\L(G)

MTPSCTLGICAPSVGGLVGGLLG reads: 4382MTPSCTLGICAPSVGGIVGGLLG reads: 0251MTPSCTLGICAPNVGGLVGGLLG reads: 0201MTPSCTMGLCVPNVGGLVGGILG reads: 0000MTPSCTMGICVPNVGGLVGGILG reads: 0000MTPSCTMGICVPNVGGLVGGLLG reads: 0000 CAPNVGG reads: 0442 CVPNVGG reads: 0000


Group 23 Fusion Toxin or Transcript Misassembly

23

looks like a fusion toxin, but could be transcript misassembly?• 12 Cys, 2 pairs of adjacent CC’s

• maybe 11 Cys in mature form, active dimer?• perhaps 2 concatenated 6 Cys toxins• Present in 2 libraries

>as:pi-theraphotoxin-Pc1a|sp:P60514... Toxin from venom of the spider Psalmopoeus cambridgei that inhibits ASIC1a channels

Score = 64.7 bits (156), Expect = 1e-13 Identities = 26/39 (66%), Positives = 28/39 (71%)

Query: 52 QECIAKWKSCAGRKLDCCEGLECWKRRWGHEVCVPITQK 90 ++CI KWK C R DCCEGLECWKRR EVCVP T KSbjct: 1 EDCIPKWKGCVNRHGDCCEGLECWKRRRSFEVCVPKTPK 39

Top BLASTHit does notspan fusion junction

pro/mature


Alignment of CCC 8 Cys containing groups 7,14,16,30

ProteinGroup

16.116.116.17.17.1

30.114.114.1

WCAKNEDCCCPMKCIGAWYN reads:312WCGKEGDCCCPWKCIGQWYN reads: 25RCNANSDCCCPLKCVIRLVG reads: 3YCEKDKDCCCPMRCVKSYWK reads: 10

Group Proposed names16.1 delta-hexatoxin-Hi3 7.1 delta-hexatoxin-Hi114.1 delta-hexatoxin-Hi230.1 delta-hexatoxin-Hi4


Coverage of CCC 8 Cys containing groups 7,14,16,30


Alignment of noCCC 8 Cys containing groups

ProteinGroup12.112.1 1.1 1.1 5.121.1 8.1 8.113.113.121.115.1 5.1 5.120.1

1,5,8,12,13,15,20,21

5,13,15,20,21ProteinGroup13.113.115.121.121.120.1 5.1 5.1 5.1

CC, CXC, CXC


Group 1

topBLAST hit not in Arachnoservergi|294653183|gb|ADF28499.1|putative mature sequence toxin-like ACSKQ [Pelinobius muticus]

<Hsp_qseq>EQIAAEENQLVEDLVQYAGTRLTQKRATRCSKKLGEKCNYHCECCGATVACSTVYVGGKETNFCSDKTSNNGALNTVGQGLNVVSNGLSAFQCWG<Hsp_midline>+ A+E ++L+E L + + Q+ A CSK++GEKC + C+CCGATV C T+YVGG C KTSNN LNT+G G+N V N ++ CWG

<Hsp_hseq>RKTASETSKLLEKL-GVSREAIPQEMARACSKQIGEKCEHDCQCCGATVVCGTIYVGGNAVEQCMSKTSNNAVLNTMGHGMNAVQNAFTSVMCWG

8 Cys

CSDKTSNNGALNTVGQGLNVVSNGLSAFQC reads: 1023

CSKKLGEKCNYHCECCGATVACS reads: 371CSKKLGEKCDYHCECCGATVACD reads: 49

CSTVYVGGKETNFC reads: 5251

TVGQGLNVVSNGLSAFQC reads: 4739

CSDKTSNNGALNTVGQGLN reads: 1887

GGKETNFCSDKTSNN reads: 3771GGRETNFCSDKTSNN reads: 10

CSKKLGEKCNYHC reads: 769CSKKLGEKCDYHC reads: 111

Evidence exists in spider.q17 reads for paralogs missing from the assembly.

N to D supported in MS/MSmatched as N to n deamidation NVVVNGFSAFQC reads: 25


Alignment of CECCG 8 Cys containing groups 1,11,29

Groups 1,11,29 are highly related.Lower level variant reads not assembled with group 1 may be the missing N-term for Group 11,29

ProteinGroup

1.1 1.111.111.129.1


Alignment of CECCG 8 Cys containing groups 1,11,12

ProteinGroup12.112.1 1.1 1.1

ProteinGroup 1.1 1.1 11.1 11.1

Group 1 and 11 are highly related.Lower level variant reads not assembled with group 1 may be the missing N-term for Group 11


Alignment of CC, CXC, CXC 8 Cys containing groups 5,13,15,20,21,26

5,13,15,20,21

CC, CXC, CXC

ProteinGroup13.113.115.121.121.120.1 5.1 5.1 5.1

ProteinGroup26.126.126.115.113.113.121.1 5.1 5.120.1


Coverage of CC, CXC, CXC 8Cys containing groups 5,20


Coverage of CC,CXC,CXC 8Cys containing groups 13,15,21,26

ProteinGroup13.113.115.121.121.1

26


Coverage of CC, CYIC 8Cys containing group 8,24

24


Alignment of 10 Cys containing protein groups 3,4,10,19

ProteinGroup 4.1 4.1 4.1 4.119.1 3.1 3.1 3.210.110.1

Groups 3,4,10 too divergent to be paralogs.Examine group 19 to determine spectral coverage of variant AAs

Group Proposed names4.1 mu1-hexatoxin-Hi1a19.1 mu1-hexatoxin-Hi1b3.1 mu1-hexatoxin-Hi2a3.2 mu1-hexatoxin-Hi2b10.1 mu1-hexatoxin-Hi3a


Coverage of 10 Cys containing groups 3,4,10,19


PSM Overlap for Group 10.1

Group 10.1 37_1680.3d.comp799_c0_seq1_F2_R4 Hi_3day_Q17_37 unknown q T C G G P D D C G E G S C C V G S F S R K C L L L S R E G A S C Q P E Y Q H T G G I/Y/S Y/A/C/P C E N/G Y I C S P I N R C Q K A 21 65 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.5243.5243.7.pkl.spo T C G G P D|D|C|G E|G|S|C|C\V\G\S\F 22 18 46 CID K120223_Hi_ri_ff4f 4f4_pccc_01.3930.3930.2.pkl.spo T C\G G P D D\C G E G S C C V G/S/F 22 18 46 HCD K120223_Hi_ri_ff4f4f4_pccc_01.3931.3931.2.pkl.spo T C/G G/P D|D C/G E/G S C C V G S F S R K 22 21 43 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4075.4075.5.pkl.spo T C/G|G P|D D\C G|E|G/S C\C V|G|S|F|S|R\K 22 21 43 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4077.4077.5.pkl.spo T C/G|G P/D\D\C\G|E\G S C\C V/G|S|F|S|R\K 22 21 43 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4064.4117.4.pkl.spo T/C|G/G|P D/D|C/G E/G S C C V/G S F S R K 22 21 43 CID K120223_Hi_ri_ff4f4f4_pccc_01.3098.3098.3.pkl.spo T C G/G P D D C G/E/G/S C C V/G/S/F/S R K 22 21 43 ETD K120223_Hi_ri_ff4f4f4_pccc_01.3100.3100.3.pkl.spo T C/G/G P D|D C G E G/S C C V G/S/F/S R\K 22 21 43 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4061.4061.3.pkl.spo T C\G G/P D D/C/G E/G/S/C/C/V/G/S/F/S R K 2 2 21 43 HCD K120223_Hi_ri_ff4f4f4_pccc_01.3099.3099.3.pkl.spo T/C/G/G/P D|D|C/G E|G S/C C V/G S F S R K C L L L 22 25 39 CID K120223_Hi_ri_ff4f4f4_pccc_01.5269.5269.3.pkl.spo T C G/G P D D C G E G S C C V G/S/F/S/R K C L L\L 22 25 39 ETD K120223_Hi_ri_ff4f4f4_pccc_01.52 71.5271.3.pkl.spo T C G G P D D C G E G S C C V G S F S R K C L L L S R E G A S C Q P E Y Q H T G G I Y S\Y/A|C/P C/E/N G Y I C S P I N R C Q K A 22 64 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.5227.5227.8.pkl.spo T C G G P D D C G E G S C C V G S F S R K C L/L L S R E G A S C Q P E Y Q H T G G I\Y\S Y A\C|P C E N/G Y I C S P I N R C Q K A 22 64 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.5292.5292.7.pkl.spo T C G G P D D\C G E G S C C V G S F S R K C L L L S R E G A S C Q P E Y Q H/T/G G I/Y S Y/A/C/P C E N/G Y I/C S/P I N R C Q K A 22 64 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.5191.5191.6.pkl.spo T C G\G P D\D C G E G S C C V G S F S R K\C L L L S R E G A S C Q P E Y Q H T G G/I Y S Y A C P C E/n G Y I C S P I n R C/Q K A 22 64 0 ETD K120223_Hi_ri_ff4f4f4_pccc_ 01.5264.5264.8.pkl.spo C/V/G/S/F/S|R\K 35 8 43 ETD K121115_Hi_dsrak_ff4f4f4C HEsa_cccc_01.3004.3121.2.pkl.spo C/V/G/S/F/S R K 35 8 43 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.3003.3120.2.pkl.spo V/G|S|F S R\K 36 7 43 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.2885.2969.2.pkl.spo V/G/S/F S/R\K 36 7 43 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.2887.2971.2.pkl.spo V/G|S|F S R/K 36 7 43 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.2886.2970.2.pkl.spo S R K C L L L S R E G A S C Q\P E Y Q H T G G I\Y\S\Y\A|C/P C/E/N G Y I C S P I N R C Q K A 40 46 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.4775.4775.7.pkl.spo S R\K\C\L\L/L\S\R\E\G\A\S\C|Q P E\Y Q\H T|G|G\I Y S Y A C P C E/N G/Y I C/S P I N/R C/Q/K A 40 46 0 ETD K120223_Hi_ri_ff4f4f4_pccc_01.4777.4777.7.pkl.spo S R K C L L L S R E G A S C Q P E Y Q\H\T\G\G\I\Y\S\Y A C/P C E N G Y I/C/S P I N R C Q K A 40 46 0 HCD K120223_Hi_ri_ff4f4f4_pccc_01.4776.4776.7.pkl.spo S R K C L L L S R E G A S C Q P E Y\Q\H/T|G G\I|Y/S/Y/A/C/P C E N/G Y/I/C S P I N R C Q K A 40 46 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.4798.4798.5.pkl.spo S R K C\L L L\S\R\E G A S C Q P E Y Q H T G\G I Y S Y A C P C E N G Y I C S P I\N R C Q K A 40 46 0 ETD K120223_Hi_ri_ff4f4f4_pccc_01.4800.4800.5.pkl.spo S R K C\L\L L S\R E G A S C Q P E Y Q H T G\G I Y S Y A C P C E N G Y I C/S P I n/R C Q K A 40 46 0 ETD K120223_Hi_ri_ff4f4f4_pccc_01.4774.4774.5.pkl.spo S R K C L L L S R E G A S C Q P E Y\Q\H\T\G/G\I\Y\S Y A C P C E n G Y/I/C S P I N R C\Q K A 40 46 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.4772.4772.5.pkl.spo C/L|L/L S R|E|G|A\S C|Q P E Y|Q H T G G I Y\S|Y A\C P C 43 28 15 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6424.6424.4.pkl.spo C L\L\L S R\E G A S C|Q P|E\Y|Q H/T G\G I Y\S\Y A C P C\E n G Y I C S P I n|R C 43 40 3 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6704.6704.7.pkl.spo C L|L/L S\R\E\G A S C|Q P\E|Y\Q H T G|G I Y S|Y A\C P C E n G Y I\C S P I\n/R C/Q\K A 43 43 0 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6494.6494.7.pkl.spo C L L L S R E G A S C Q\P E Y Q H T G G I\Y\S\Y\A\C\P C E n G Y I C S P I n R C Q K A 43 43 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.5246.5246.5.pkl.spo C L\L L S\R\E\G A S C Q P\E Y Q H T G G I Y S Y A C P C E n G Y I C S P I n/R C/Q K A 43 43 0 ETD K120223_Hi_ri_ff4f4f4_pccc_01.5248.5248.5.pkl.spo S R E G A S C Q P E Y Q H T G G I\Y\S\Y|A|C|P C E N G Y I C S P I N R C Q K A 47 39 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.4752.4752.5.pkl.spo S Y/A|C|P/C/E/N/G Y/I/C S P I N R C Q K A 65 21 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.4044.4099.3.pkl.spo S Y\A\C P C E N G Y I C S P I/N R C Q K A 65 21 0 HCD K120223_Hi_ri_ff4f4f4_pccc_01.4045.4100.3.pkl.spo A C|P/C/E/N/G Y I/C S P I N R C Q K A 67 19 0 CID K120223_Hi_ri_ff4f4f4_pccc_01.3631.3680.3.pkl.spo A C P C E|N/G|Y/I C|S P I N/R C/Q K A 67 19 0 ETD K120223_Hi_ri_ff4f4f4_pccc_01.3633.3682.3.pkl.spo A C|P C E N G Y/I/C/S/P I/N R C Q K A 67 19 0 HCD K120223_Hi_ri_ff4f4f4_pccc_01.3632.3632.3.pkl.spo E N/G/Y I/C/S/P I N R C Q K A 71 15 0 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4825.4825.4.pkl.spo E/N\G|Y\I/C|S P I|N|R\C|Q|K A 71 15 0 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4827.4827.4.pkl.spo E N/G Y I/C/S/P I N R C Q K A 71 15 0 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4826.4826.4.pkl.spo E/N/G/Y/I/C/S/P I N R C Q K A 71 15 0 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4819.4849.3.pkl.spo E/N G/Y/I C|S P I|N|R C|Q\K\A 71 15 0 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4821.4851.3.pkl.spo E/N/G\Y/I/C/S P I N R C Q K A 71 15 0 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4820.4850.3.pkl.spo S/P|I/N R C Q K\A 77 9 0 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.2469.2469.3.pkl.spo S P/I|N|R\C|Q|K A 77 9 0 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.2471.2471.3.pkl.spo S/P|I/N R C Q K\A 77 9 0 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.2470.2470.3.pkl.spo S P|I|N R\C Q|K A 77 9 0 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.2460.2460.2.pkl.spo


Coverage of 6Cys containing groups 6, 9


Coverage of 6Cys groups 17, 22, 25, 27, 28

27

22

25

28

17


Group 18 - 6Cys - only intact MS/MS z5-10

(R)M F C K P L D\Q|Q\C\N K|D L H\C C|K P L/K C/R|R/S/N/N/G/R/K Y C/K P(-) (R)M F\C\K P L D\Q|Q\C N K\D L H\C\C|K P/L/K C/R/R/S/N/N/G R/K Y C/K P(-) (R)M F\C\K P L D\Q|Q\C N K\D L H\C C K P L K C|R/R/S/N/N/G/R/K/Y C/K P(-) (R)M F C\K P L D\Q\Q\C N K\D/L H\C\C|K P L K\C/R/R S/N/N/G/R/K/Y C K P(-) (R)M/F C/K P L\D\Q|Q\C N K D L H C C K P L K\C\R\R/S/N/N/G/R/K Y C K P(-)

z9z8z7z6z5

(R)M/F/C\K/P L D\Q Q C N K D|L H/C C K P L K C R R S N N G R K\Y C K\P(-) z6z5(R)M/F/C|K/P L D\Q Q/C N/K/D|L H C C K P L K C R R S N N G R K\Y C K\P(-)

(R)M F C|K/P/L/D/Q/Q/C/N/K/D/L H/C C K P L K C R R S N N G R K\Y C K P(-) z5

ETD

CID

HCD

CysIodoacetamide

56spectra

2peptides


Group 18 - 6Cys - only intact MS/MS z5-10

CysIodoacetamide

56spectra

2peptides

MFCKPLDQQCN reads: 6

CRRSNNGRKYCKP reads: 12

CNKDLHCCKPLKC reads: 1

CCKPLKCRR reads: 11CRKPLKKRL reads: 66


Group 18 - 6Cys - only intact MS/MS z8-9 ETD(R)M F C K P L D\Q|Q\C\N K|D L H\C C|K P L/K C/R|R/S/N/N/G/R/K Y C/K P(-)

(R)M F\C\K P L D\Q|Q\C N K\D L H\C\C|K P/L/K C/R/R/S/N/N/G R/K Y C/K P(-)


Group 18 - 6Cys - only intact MS/MS z6-7, ETD(R)M F\C\K P L D\Q|Q\C N K\D L H\C C K P L K C|R/R/S/N/N/G/R/K/Y C/K P(-)

(R)M F C\K P L D\Q\Q\C N K\D/L H\C\C|K P L K\C/R/R S/N/N/G/R/K/Y C K P(-)


Group 18 - 6Cys - only intact MS/MS z5(R)M/F C/K P L\D\Q|Q\C N K D L H C C K P L K\C\R\R/S/N/N/G/R/K Y C K P(-)

(R)M F C|K/P/L/D/Q/Q/C/N/K/D/L H/C C K P L K C R R S N N G R K\Y C K P(-)

ETD

HCD


BLAST2GO – Functional Annotation

• Export FASTA of Valid Hits from SM• Run BLAST step• Run GO mapping step• Run annotation step

• Run InterProScan, SIGNALP• Run GO-Slim• Export results to SM categories file


Venom Toxin Nomenclature

King GF, Gentz MC, Escoubas P, Nicholson GM A rational nomenclature for naming peptide toxins from spiders and other venomous animals.Toxicon 52 (2008) 264–276.


Next Steps

• Improve paralog inclusion in RNA-Seq Transcript Assembly.• Call full length gene, predict signal & propeptide.

• 10 Cys toxins lack propeptide?• CCC 8 cys toxins propeptide so long that the assembly

doesn’t extend upstream enough to cover signal peptide?• Group 23 looks like a fusion toxin, misassembled?

• 12 Cys, 2 pairs of adjacent CC’s• perhaps 2 concatenated 6 Cys toxins• Present in 2 libraries

• Run SM homology searches• Improve SM monoisotopic m/z assignment for z>4.• Obtain ETD MS/MS of intact toxins after MeAziridine Cys

mod.• Assemble spectra de novo.• Name novel toxins.


Adding Charge to Cys for better ETD

Fig. 1. Overview of the de novo sequencing strategy. (I) UV trace of HPLCseparation of crude venom extract from C. textile. (II) MALDI TOF MS offraction i after no treatment, reduction, and alkylation. (III) On-line LC ESIMS/MS using CAD and ETD on reduced and alkylated aliquots of fraction i. The final step shows the conversion of Cys residues to dimethylated Lys analogs followed by ETD fragmentation; MS/MS is shown for the (M5H)5 ion of the 1,889.715 Da species in II. c ions are indicated by and z ions by . Shell image Copyright 2005, Richard Ling.

Ueberheide BM, Fenyo¨ D, Alewood PF, and Chait BT. PNAS 2009 106 , 6910–6915 .

2-Methylaziridine


Lys-C Cleaves at MeAziridine Cys mod

Group 4.1 u1-hexatoxin-hi1a 37_1680.3d.comp452_c0_seq2_F2_R3 Hi_3day_Q17_37 C G|D|Q|V C|G\A|G|T\C 37 11 59 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.0830.0857.3.pkl.spo C G|D|Q|V C|G|A|G|T\C 37 11 59 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.2261.2381.3.pkl.spo C G/D/Q/V C G\A G\T\C 37 11 59 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.0810.0947.2.pkl.spo C G/D/Q|V C\G\A\G\T\C 37 11 59 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.2177.2361.2.pkl.spo C G/D/Q/V C G\A\G\T\C 37 11 59 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.2391.2427.2.pkl.spo C G|D|Q V\C G|A\G|T\C\C|A/E|Y P|E I\H\C\K 37 21 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4477.4477.6.pkl.spo C G D Q V C G A G T C C/A E/Y/P E I H C K 37 21 49 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4476.4476.6.pkl.spo C|G|D|Q V\C|G|A G\T|C/C A E|Y P|E|I|H\C\K 37 21 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4381.4431.5.pkl.spo C|G|D|Q V\C\G|A G\T\C|C|A\E|Y P|E|I|H\C\K 37 21 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4491.4491.5.pkl.spo C/G|D|Q\V\C|G|A G\T|C|C\A E|Y P|E|I|H\C\K 37 21 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4531.4531.5.pkl.spo C/G|D|Q V C|G/A\G\T|C\C\A|E|Y P|E|I|H\C\K 37 21 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4411.4461.4.pkl.spo C G D/Q V/C G A G T C C/A E/Y|P E\I\H\C\K 37 21 49 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4522.4522.3.pkl.spo C/G/D/Q V C\G\A G|T C C A E|Y P|E|I/H\C\K 37 21 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4387.4441.3.pkl.spo C/G/D/Q V C|G A G/T C C A E|Y P|E/I/H\C\K 37 21 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4471.4481.3.pkl.spo G A/G T C C A E/Y/P E|I H C K 43 15 49 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4225.4299.4.pkl.spo G/A/G/T\C|C|A|E|Y P|E|I\H|C\K 43 15 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4227.4301.4.pkl.spo G A\G T C C A E/Y/P E I H C K 43 15 49 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4226.4300.4.pkl.spo G A/G T C C A E|Y|P E\I H\C\K 43 15 49 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4259.4259.3.pkl.spo G/A/G/T C\C|A|E|Y P|E|I\H\C\K 43 15 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4261.4304.3.pkl.spo G A/G T C/C/A E|Y|P E\I\H\C\K 43 15 49 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4292.4292.2.pkl.spo G A G/T C C/A/E/Y P\E\I H C\K 43 15 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4294.4294.2.pkl.spo G A/G T C/C/A/E|Y|P E I|H|C\K 43 15 49 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4293.4293.2.pkl.spo C/A|E|Y/P/E|I\H|C\K 48 10 49 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4134.4237.3.pkl.spo R V\G\Q|L|Y|D|I C V D S E A T K 58 16 33 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6342.6489.4.pkl.spo R/V|G|Q/L|Y/D|I\C/V|D|S|E|A|T\K 58 16 33 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6344.6491.4.pkl.spo R V\G\Q\L\Y/D/I/C/V D/S E/A/T K 58 16 33 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6343.6490.4.pkl.spo R V G Q\L\Y\D|I|C/V/D|S E\A T\K 58 16 33 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6329.6526.3.pkl.spo R/V/G|Q|L|Y/D|I\C|V|D|S|E|A\T\K 58 16 33 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6331.6528.3.pkl.spo R V G Q\L\Y\D\I|C/V/D/S E A/T/K 58 16 33 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6330.6527.3.pkl.spo R V G Q L|Y|D|I\C|V|D|S E\A\T\K 58 16 33 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6336.6486.2.pkl.spo R/V/G/Q L\Y\D I C\V\D\S\E\A\T\K 58 16 33 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6338.6488.2.pkl.spo R V G Q L\Y\D|I\C V|D|S E|A|T|K 58 16 33 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6337.6487.2.pkl.spo L Y/D|I|C/V D|S E A T K 62 12 33 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6372.6416.2.pkl.spo L/Y/D/I C/V|D|S\E\A\T\K 62 12 33 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6374.6418.2.pkl.spo L Y|D|I|C/V D/S E/A/T/K 62 12 33 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6373.6417.2.pkl.spo Y/D|I|C/V D|S/E/A/T K 63 11 33 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6346.6496.2.pkl.spo Y/D/I C/V D|S|E\A\T\K 63 11 33 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6348.6498.2.pkl.spo Y D|I|C/V D/S/E/A/T/K 63 11 33 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6347.6497.2.pkl.spo D/I|C/V/D|S E/A\T K 64 10 33 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6359.6469.2.pkl.spo D/I C/V/D/S|E\A\T\K 64 10 33 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6361.6471.2.pkl.spo D I|C/V/D/S E/A/T/K 64 10 33 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6360.6470.2.pkl.spo D/S/G|N/H/L|F\F\C P C|D|E|G|m|Y\C 74 17 16 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6534.6591.4.pkl.spo D/S/G/N/H\L|F|F C P\C|D|E\G|m\Y\C 74 17 16 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6514.6514.3.pkl.spo D/S/G/N/H|L|F/F\C P\C|D|E/G|m\Y\C 74 17 16 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6548.6588.3.pkl.spo D/S/G/N H|L|F\F\C P C D E G M\Y\C|D|m\N/S W S\C 74 24 9 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.7231.7231.4.pkl.spo D S G N H\L F|F\C P C|D|E|G\m Y\C|D/m N|S W\S\C 74 24 9 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6944.6944.5.pkl.spo D/S G N H L F F C/P C D E G m\Y C D m\N\S\W\S\C 74 24 9 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6936.6936.4.pkl.spo D/S/G/N H\L|F\F\C P C\D\E\G\m Y C|D|m|N|S W\S\C 74 24 9 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6938.6938.4.pkl.spo D/S G/N H|L\F F\C P C\D\E\G\m Y\C\D|m\n/S W S C 74 24 9 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6968.6968.4.pkl.spo D/S G N H|L F/F\C P\C|D|E\G\m|Y\C|D|m|N|S W|S\C/Q\K 74 26 7 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6581.6581.6.pkl.spo D S\G N H\L\F F C P C D E G m Y C D m N S/W/S C Q K 74 26 7 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6580.6580.6.pkl.spo D S G N H\L F\F\C P C\D|E\G\m Y C|D|m|N\S\W/S\C|Q\K 74 26 7 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6541.6541.5.pkl.spo D S/G/N H|L F|F\C P C\D\E\G\m Y\C|D|m|N\S\W\S\C|Q\K 74 26 7 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6578.6578.5.pkl.spo D S\G N H L\F F\C P C D E G m/Y C D m N\S|W/S C Q K 74 26 7 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6540.6540.5.pkl.spo D S|G N H L F F C/P C D E G m Y C D m N\S|W/S C Q K 74 26 7 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6577.6577.5.pkl.spo D/S/G/N H\L|F F C P C D E G m Y C|D m\N|S\W|S\C\Q\K 74 26 7 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6538.6538.4.pkl.spo D/S G N H|L F\F\C P\C|D|E|G\m\Y\C\D\m\n|S W|S\C/Q\K 74 26 7 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6574.6574.6.pkl.spo D S\G N H L\F F C P C D E G m Y C D m n S/W/S C Q K 74 26 7 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6573.6573.6.pkl.spo D/S G N H|L F\F\C P C\D|E|G\m Y\C|D\m\n S|W/S C Q K 74 26 7 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6604.6604.5.pkl.spo D S\G n H L F/F C P C D E G m Y C D m N S/W/S C Q K 74 26 7 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6603.6603.5.pkl.spo D/S G N/H|L F|F\C P C D|E\G\m\Y\C|D m/N\S W\S C|Q|K K 74 27 6 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6228.6228.5.pkl.spo D S\G N H L F\F\C P C D E G m Y C D m N S\W/S C Q K K 74 27 6 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6227.6227.5.pkl.spo D S\G n H L F F C P C D E G m Y C D m n S W/S C Q K/K 74 27 6 HCD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.6233.6233.6.pkl.spo D m|N|S\W|S|C 91 7 9 CID K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4175.4175.2.pkl.spo D/m/N/S W\S\C 91 7 9 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.4177.4177.2.pkl.spo D/m|N|S\W\S\C|Q|K\K 91 10 6 ETD K121115_Hi_dsrak_ff4f4f4CHEsa_cccc_01.3327.3327.3.pkl.spo

Cys cleavages observed only in most abundant proteinsKinetics: K >> C


MeAziridine Cys mod Yields Great ETD Spectra(K)S/Y\W|K/G|H/G\V\C\S A\S|L|F|E|R|L|K\G\C(-)

(K)C/I|G|Q|W|Y/N|G|Q|A S C|Q|S/T\F|m|G|L\F\K(S)


MeAziridine Cys mod Yields Great ETD Spectra(K)C/N|Y|H\C|E\C\C|G/A T V\A\C|S/T|V\Y/V|G G\K(E)

(K)C Y\C/D|Y|G|L|F|G|N\C|N\C|Y\K(R)


MeAziridine Cys mod Yields Great ETD Spectra

(Q)T C/G|G P|D D\C G|E|G/S C\C V|G|S|F|S|R\K(C)

(N)W C|A|K/N\E|D\C\C\C P M\K(C)

Karl ClauserProteomics and Biomarker Discovery

CentipedesSpidersScorpions

S. Morsitans

A. xerolimniorum

L. weigensis

H. jugulans

C. westwoodi

E. rubripes

U. manicatus

L. variatus

L. buchari

C. squama

I. vescus

Thereuopoda sp.

S. foelschei

C. Tropix

H. infensa

7 scorpions4 centipedes1 spider

Documents

Karl Clauser Proteomics and Biomarker Discovery Sample Experiment Summary 1 Spider 1 ug venom Reduce/alkylate De-salt LC-MS/MS 300ng C18 300A Microsorb