IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ An intracellular serpin regulates necrosis by inhibiting the induction and sequelae of lysosomal injury. An intracellular serpin regulates necrosis by inhibiting the induction and sequelae of lysosomal injury. Cell (0092-8674) 2007 imex curation Luke CJ., Pak SC., Askew YS., Naviglia TL., Askew DJ., Nobar SM., Vetica AC., Long OS., Watkins SC., Stolz DB., Barstead RJ., Moulder GL., Bromme D., Silverman GA. To detect SRP-6 peptidase inhibitor activity, different peptidases and a 10-25-fold molar excess SRP-6 were incubated for 30 min at 25 C in the appropriate reaction buffer. Residual enzyme activity was measured by addition of substrate. Activity of enzyme in the presence of inhibitor was compared to an uninhibited control. CatV (L2) is stated to be of human origin in the cited reference Bromme et al. (1999) (PMID 10029531). CatK (O2) and CatL are stated to be of human origin in the cited reference Bromme et al. (1996) (PMID 8567669). The species of origin of a number of the peptidases in Table S1 could not be determined and thus this data has not been added. gsilverman@mail.magee.edu 01-Feb-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 6 in vitro In vitro enzymatic study enzymatic study experimental particp experimental participant identification experimental particp Luke CJ., Pak SC., Askew YS., Naviglia TL., Askew DJ., Nobar SM., Vetica AC., Long OS., Watkins SC., Stolz DB., Barstead RJ., Moulder GL., Bromme D., Silverman GA. To detect SRP-6 peptidase inhibitor activity, different peptidases and a 10-25-fold molar excess SRP-6 were incubated for 30 min at 25 C in the appropriate reaction buffer. Residual enzyme activity was measured by addition of substrate. Activity of enzyme in the presence of inhibitor was compared to an uninhibited control. CatV (L2) is stated to be of human origin in the cited reference Bromme et al. (1999) (PMID 10029531). CatK (O2) and CatL are stated to be of human origin in the cited reference Bromme et al. (1996) (PMID 8567669). The species of origin of a number of the peptidases in Table S1 could not be determined and thus this data has not been added. gsilverman@mail.magee.edu Accepted 2008-FEB-9 by DAVET. 01-Feb-2008: Contacted by JYOTI. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 6 imex curation An intracellular serpin regulates necrosis by inhibiting the induction and sequelae of lysosomal injury. An intracellular serpin regulates necrosis by inhibiting the induction and sequelae of lysosomal injury. Cell (0092-8674) 2007 imex curation Luke CJ., Pak SC., Askew YS., Naviglia TL., Askew DJ., Nobar SM., Vetica AC., Long OS., Watkins SC., Stolz DB., Barstead RJ., Moulder GL., Bromme D., Silverman GA. To detect SRP-6 peptidase inhibitor activity, different peptidases and a 10-25-fold molar excess SRP-6 were incubated for 30 min at 25 C in the appropriate reaction buffer. Residual enzyme activity was measured by addition of substrate. Activity of enzyme in the presence of inhibitor was compared to an uninhibited control. CatV (L2) is stated to be of human origin in the cited reference Bromme et al. (1999) (PMID 10029531). CatK (O2) and CatL are stated to be of human origin in the cited reference Bromme et al. (1996) (PMID 8567669). The species of origin of a number of the peptidases in Table S1 could not be determined and thus this data has not been added. gsilverman@mail.magee.edu 01-Feb-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 6 in vitro In vitro comigration in sds comigration in sds page comigration in sds weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Luke CJ., Pak SC., Askew YS., Naviglia TL., Askew DJ., Nobar SM., Vetica AC., Long OS., Watkins SC., Stolz DB., Barstead RJ., Moulder GL., Bromme D., Silverman GA. To detect SRP-6 peptidase inhibitor activity, different peptidases and a 10-25-fold molar excess SRP-6 were incubated for 30 min at 25 C in the appropriate reaction buffer. Residual enzyme activity was measured by addition of substrate. Activity of enzyme in the presence of inhibitor was compared to an uninhibited control. CatV (L2) is stated to be of human origin in the cited reference Bromme et al. (1999) (PMID 10029531). CatK (O2) and CatL are stated to be of human origin in the cited reference Bromme et al. (1996) (PMID 8567669). The species of origin of a number of the peptidases in Table S1 could not be determined and thus this data has not been added. gsilverman@mail.magee.edu Accepted 2008-FEB-9 by DAVET. 01-Feb-2008: Contacted by JYOTI. Cell (0092-8674) 2007 DAVET Only protein-protein interactions 6 imex curation catl2_human Cathepsin L2 CTSV CATL2 CTSL2 CTSU Cathepsin U Cathepsin V UNQ268/PRO305 protein protein human Homo sapiens Human MNLSLVLAAFCLGIASAVPKFDQNLDTKWYQWKATHRRLYGANEEGWRRAVWEKNMKMIELHNGEYSQGKHGFTMAMNAFGDMTNEEFRQMMGCFRNQKFRKGKVFREPLFLDLPKSVDWRKKGYVTPVKNQKQCGSCWAFSATGALEGQMFRKTGKLVSLSEQNLVDCSRPQGNQGCNGGFMARAFQYVKENGGLDSEESYPYVAVDEICKYRPENSVANDTGFTVVAPGKEKALMKAVATVGPISVAMDAGHSSFQFYKSGIYFEPDCSSKNLDHGVLVVGYGFEGANSNNSKYWLVKNSWGPEWGSNGYVKIAKDKNNHCGIATAASYPNV CD2DE51AC5F242C8 catk_human Cathepsin K Cathepsin O Cathepsin X Cathepsin O2 CTSO CTSO2 CTSK protein protein human Homo sapiens Human MWGLKVLLLPVVSFALYPEEILDTHWELWKKTHRKQYNNKVDEISRRLIWEKNLKYISIHNLEASLGVHTYELAMNHLGDMTSEEVVQKMTGLKVPLSHSRSNDTLYIPEWEGRAPDSVDYRKKGYVTPVKNQGQCGSCWAFSSVGALEGQLKKKTGKLLNLSPQNLVDCVSENDGCGGGYMTNAFQYVQKNRGIDSEDAYPYVGQEESCMYNPTGKAAKCRGYREIPEGNEKALKRAVARVGPVSVAIDASLTSFQFYSKGVYYDESCNSDNLNHAVLAVGYGIQKGNKHWIIKNSWGENWGNKGYILMARNKNNACGIANLASFPKM DIP protein P43235 original sequence version: 121 4677C3C89FF4CE85 can2_rat Calpain-2 catalytic subunit Capn2 Calpain-2 large subunit Calcium-activated neutral proteinase 2 Calpain M-type Millimolar-calpain protein protein rat Rattus norvegicus (Rat) Rat MAGIAMKLAKDREAAEGLGSHERAIKYLNQDYETLRNECLEAGALFQDPSFPALPSSLGFKELGPYSSKTRGIEWKRPTEICADPQFIIGGATRTDICQGALGDCWLLAAIASLTLNEEILARVVPLDQSFQENYAGIFHFQFWQYGEWVEVVVDDRLPTKDGELLFVHSAEGSEFWSALLEKAYAKINGCYEALSGGATTEGFEDFTGGIAEWYELRKPPPNLFKIIQKALEKGSLLGCSIDITSAADSEAVTYQKLVKGHAYSVTGAEEVESSGSLQKLIRIRNPWGQVEWTGKWNDNCPSWNTVDPEVRANLTERQEDGEFWMSFSDFLRHYSRLEICNLTPDTLTCDSYKKWKLTKMDGNWRRGSTAGGCRNYPNTFWMNPQYLIKLEEEDEDDEDGERGCTFLVGLIQKHRRRQRKMGEDMHTIGFGIYEVPEELTGQTNIHLSKNFFLTTRARERSDTFINLREVLNRFKLPPGEYVLVPSTFEPHKNGDFCIRVFSEKKADYQTVDDEIEANIEEIEANEEDIGDGFRRLFAQLAGEDAEISAFELQTILRRVLAKREDIKSDGFSIETCKIMVDMLDEDGSGKLGLKEFYILWTKIQKYQKIYREIDVDRSGTMNSYEMRKALEEAGFKLPCQLHQVIVARFADDELIIDFDNFVRCLVRLEILFKIFKQLDPENTGTIQLDLISWLSFSVL DIP protein Q07009 original sequence version: 129 296B0DC3BEEF5B90 catl1_human Procathepsin L CTSL CTSL1 Major excreted protein Cathepsin L1 protein protein human Homo sapiens Human MNPTLILAAFCLGIASATLTFDHSLEAQWTKWKAMHNRLYGMNEEGWRRAVWEKNMKMIELHNQEYREGKHSFTMAMNAFGDMTSEEFRQVMNGFQNRKPRKGKVFQEPLFYEAPRSVDWREKGYVTPVKNQGQCGSCWAFSATGALEGQMFRKTGRLISLSEQNLVDCSGPQGNEGCNGGLMDYAFQYVQDNGGLDSEESYPYEATEESCKYNPKYSVANDTGFVDIPKQEKALMKAVATVGPISVAIDAGHESFLFYKEGIYFEPDCSSEDMDHGVLVVGYGFESTESDNNKYWLVKNSWGEEWGMGGYVKMAKDRRNHCGIASAASYPTV 8CD17D00EF859D85 g5efh4_caeel srp-6 CELE_C03G6.19 C03G6.19 protein protein caeel Caenorhabditis elegans MSDSSSDEKMGLLLHSETDFGLSLLRQQNLTESFVFSPLSIALALSLVHVAAKGETRDEIRKALLNGATDEELEQHFSNISAGLLVAEKGTEVNVANHIFSRKTFTIKKLYLNDVKKLYNAGASQLNFEDQEASAEAINNFVSENTKGHIKKIINPDSISEELVAVLTNAFYFKANWQTKFKKESTYKREFFSSENSKRETEFLHSRNSNRKYSENGQFQVLSLPYKDTSFALSIFLPKTRFGLSEALQNLDSVTIQQLMSNTSNTLVNIAMPKWKIETALGLNRALMAVGIEKAFTDSADLSNFADGIYISQAAHKALIEVDEDGTVAAAATTISFSLTSVFIPAEEPIEFTADHPFLFILSKDNHPLFIGIHN BEFE7D31BEF0275E srp_6-ctsk 1 4 enzyme enzyme unspecified role unspecified role spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 7 inhibitor inhibitor unspecified role unspecified role physical association physical association Table S1 recombinant SRP-6 inhibited papain-like lysosomal cysteine peptidases (clan CA, family C1) such as catL and catK, Inhibition (%) = 94 ([I]0/[E]0 = 25) srp_6-ctsl2 1 7 inhibitor inhibitor unspecified role unspecified role 3 enzyme enzyme unspecified role unspecified role picpa Pichia pastoris (Yeast) Yeast Pichia pastoris physical association physical association Table S1 Inhibition (%) = 99 ([I]0/[E]0 = 25) srp_6-ctsl 1 7 inhibitor inhibitor unspecified role unspecified role 6 enzyme enzyme unspecified role unspecified role physical association physical association 4 A, C, Table S1 Inhibition (%) = 99 ([I]0/[E]0 = 25); complete inhibition at inhibitor:enzyme ratio of approx. 1:1; second-order rate constant ka = 10 E4 +- 0.1 M-1 s-1 (mean ka +- SD from 3-4 experiments) recombinant SRP-6 inhibited papain-like lysosomal cysteine peptidases (clan CA, family C1) such as catL srp_6-capn2 1 7 inhibitor inhibitor unspecified role unspecified role 5 enzyme enzyme unspecified role unspecified role physical association physical association 4 B, D, Table S1 Inhibition (%) = 96 ([I]0/[E]0 = 5); complete inhibition at inhibitor:enzyme ratio of approx. 1:1.5; second-order rate constant ka = 2.3 E4 +- O.4 M-1 s-1 (mean ka +- SD from 3-4 experiments) The stoichiometry of inhibition (SI) for the interaction of SRP-6 with catL (A) or calpain-2 (B). Inhibitor:enzyme ratio of 1:1 and 1:1.5 completely inhibited catL and calpain-2 activity, respectively. srp_6-ctsl-1 2 6 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction 4 E Autoradiogram after SDS-PAGE revealed native or cleaved SRP-6 (*) and a higher molecular mass complex with catL or calpain-2 srp_6-capn2-1 2 5 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction 4 F Autoradiogram after SDS-PAGE revealed native or cleaved SRP-6 (*) and a higher molecular mass complex with catL or calpain-2