ID CYSP_TRYCR Reviewed; 467 AA. AC P25779; DT 01-MAY-1992, integrated into UniProtKB/Swiss-Prot. DT 01-MAY-1992, sequence version 1. DT 27-MAR-2024, entry version 144. DE RecName: Full=Cruzipain; DE EC=3.4.22.51; DE AltName: Full=Cruzaine; DE AltName: Full=Major cysteine proteinase; DE Flags: Precursor; OS Trypanosoma cruzi. OC Eukaryota; Discoba; Euglenozoa; Kinetoplastea; Metakinetoplastina; OC Trypanosomatida; Trypanosomatidae; Trypanosoma; Schizotrypanum. OX NCBI_TaxID=5693; RN [1] RP NUCLEOTIDE SEQUENCE. RC STRAIN=Tulahuen; RX PubMed=1559982; DOI=10.1016/s0021-9258(18)42533-1; RA Eakin A.E., Mills A.A., Harth G., McKerrow J.H., Craik C.S.; RT "The sequence, organization, and expression of the major cysteine protease RT (cruzain) from Trypanosoma cruzi."; RL J. Biol. Chem. 267:7411-7420(1992). RN [2] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] (CLONES 1800-2 AND 1800-4). RC STRAIN=Tulahuen 2; RX PubMed=1311053; DOI=10.1016/0166-6851(92)90219-a; RA Campetella O., Henriksson J., Aaslund L., Frasch A.C.C., Pettersson U., RA Cazzulo J.J.; RT "The major cysteine proteinase (cruzipain) from Trypanosoma cruzi is RT encoded by multiple polymorphic tandemly organized genes located on RT different chromosomes."; RL Mol. Biochem. Parasitol. 50:225-234(1992). RN [3] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 141-306. RC STRAIN=RA; RX PubMed=2406590; DOI=10.1016/0166-6851(90)90002-4; RA Eakin A.E., Bouvier J., Sakanari J.A., Craik C.S., McKerrow J.H.; RT "Amplification and sequencing of genomic DNA fragments encoding cysteine RT proteases from protozoan parasites."; RL Mol. Biochem. Parasitol. 39:1-8(1990). RN [4] RP NUCLEOTIDE SEQUENCE [MRNA] OF 295-467. RC STRAIN=Tulahuen 2; RX PubMed=2038364; DOI=10.1016/0166-6851(91)90103-d; RA Aaslund L., Henriksson J., Campetella O., Frasch A.C.C., Pettersson U., RA Cazzulo J.J.; RT "The C-terminal extension of the major cysteine proteinase (cruzipain) from RT Trypanosoma cruzi."; RL Mol. Biochem. Parasitol. 45:345-348(1991). RN [5] RP AUTOCATALYSIS OF C-TERMINAL. RC STRAIN=Tulahuen 2; RX PubMed=2011151; DOI=10.1016/0166-6851(91)90216-s; RA Hellman U., Wernstedt C., Cazzulo J.J.; RT "Self-proteolysis of the cysteine proteinase, cruzipain, from Trypanosoma RT cruzi gives a major fragment corresponding to its carboxy-terminal RT domain."; RL Mol. Biochem. Parasitol. 44:15-21(1991). RN [6] RP SPECIFICITY. RC STRAIN=Tulahuen 2; RX PubMed=2407295; DOI=10.1016/0167-4838(90)90166-d; RA Cazzulo J.J., Cazzulo-Franke M.C., Martinez J., Franke de Cazzulo B.M.; RT "Some kinetic properties of a cysteine proteinase (cruzipain) from RT Trypanosoma cruzi."; RL Biochim. Biophys. Acta 1037:186-191(1990). RN [7] RP PROTEIN SEQUENCE OF 123-146 AND 304-317. RX PubMed=2651912; DOI=10.1016/0166-6851(89)90039-x; RA Cazzulo J.J., Couso R., Raimondi A., Wernstedt C., Hellman U.; RT "Further characterization and partial amino acid sequence of a cysteine RT proteinase from Trypanosoma cruzi."; RL Mol. Biochem. Parasitol. 33:33-42(1989). RN [8] RP X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF 123-337. RX PubMed=9260273; DOI=10.1002/pro.5560060801; RA Gillmor S.A., Craik C.S., Fletterick R.J.; RT "Structural determinants of specificity in the cysteine protease cruzain."; RL Protein Sci. 6:1603-1611(1997). CC -!- FUNCTION: Hydrolyzes chromogenic peptides at the carboxyl Arg or Lys; CC requires at least one more amino acid, preferably Arg, Phe, Val or Leu, CC between the terminal Arg or Lys and the amino-blocking group. CC -!- FUNCTION: The cysteine protease may play an important role in the CC development and differentiation of the parasites at several stages of CC their life cycle. CC -!- CATALYTIC ACTIVITY: CC Reaction=Broad endopeptidase specificity similar to that of cathepsin CC L.; EC=3.4.22.51; CC -!- ACTIVITY REGULATION: Strongly inhibited by E-64 (L-trans- CC epoxysuccinylleucylamido(4-guanidino)butane), Leupeptin, and N-alpha-p- CC tosyl-L-lysine chloromethyl ketone. CC -!- DEVELOPMENTAL STAGE: Present in all developmental stages. CC -!- MISCELLANEOUS: Purified cruzipain is able to degrade itself, yielding a CC complex mixture of small peptides, and a major 25 kDa fragment. CC -!- SIMILARITY: Belongs to the peptidase C1 family. {ECO:0000255|PROSITE- CC ProRule:PRU10088, ECO:0000255|PROSITE-ProRule:PRU10089, CC ECO:0000255|PROSITE-ProRule:PRU10090}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; M84342; AAA30181.1; -; Genomic_DNA. DR EMBL; M69121; AAA30269.1; -; Genomic_DNA. DR EMBL; M69121; AAA30270.1; -; Genomic_DNA. DR EMBL; X54414; CAA38278.1; -; mRNA. DR EMBL; M27305; AAA30180.1; -; Genomic_DNA. DR PIR; A60667; A60667. DR PIR; S16162; S16162. DR PDB; 1AIM; X-ray; 2.00 A; A=123-337. DR PDB; 1EWL; X-ray; 2.00 A; A=123-337. DR PDB; 1EWM; X-ray; 2.00 A; A=123-337. DR PDB; 1EWO; X-ray; 2.10 A; A=123-337. DR PDB; 1EWP; X-ray; 1.75 A; A=123-337. DR PDB; 1F29; X-ray; 2.15 A; A/B/C=123-337. DR PDB; 1F2A; X-ray; 1.60 A; A=123-337. DR PDB; 1F2B; X-ray; 1.80 A; A=123-337. DR PDB; 1F2C; X-ray; 2.00 A; A=123-337. DR PDB; 1ME3; X-ray; 1.20 A; A=123-337. DR PDB; 1ME4; X-ray; 1.20 A; A=123-337. DR PDB; 1U9Q; X-ray; 2.30 A; X=123-337. DR PDB; 2AIM; X-ray; 2.20 A; A=123-337. DR PDB; 2OZ2; X-ray; 1.95 A; A/C=123-337. DR PDB; 3HD3; X-ray; 1.75 A; A/B=123-337. DR PDB; 3I06; X-ray; 1.10 A; A=123-337. DR PDB; 3IUT; X-ray; 1.20 A; A=123-337. DR PDB; 3KKU; X-ray; 1.28 A; A=123-337. DR PDB; 3LXS; X-ray; 1.50 A; A/C=123-337. DR PDB; 4KLB; X-ray; 2.62 A; A/B/C/D/E=123-337. DR PDB; 4PI3; X-ray; 1.27 A; A/B=122-337. DR PDB; 4QH6; X-ray; 3.13 A; A/B/C/D/E=123-337. DR PDB; 4W5B; X-ray; 2.70 A; A/B/C=123-337. DR PDB; 4W5C; X-ray; 3.27 A; A/B/C/D/E=122-337. DR PDB; 4XUI; X-ray; 2.51 A; A/B/C=122-337. DR PDB; 6N3S; X-ray; 1.19 A; A/B=123-337. DR PDB; 6O2X; X-ray; 1.19 A; A/B=123-337. DR PDB; 6UX6; X-ray; 1.94 A; A=123-337. DR PDB; 7JUJ; X-ray; 2.20 A; A/B/C/D/E/F=123-337. DR PDB; 7S18; X-ray; 2.14 A; A=123-337. DR PDB; 7S19; X-ray; 2.08 A; A=123-337. DR PDBsum; 1AIM; -. DR PDBsum; 1EWL; -. DR PDBsum; 1EWM; -. DR PDBsum; 1EWO; -. DR PDBsum; 1EWP; -. DR PDBsum; 1F29; -. DR PDBsum; 1F2A; -. DR PDBsum; 1F2B; -. DR PDBsum; 1F2C; -. DR PDBsum; 1ME3; -. DR PDBsum; 1ME4; -. DR PDBsum; 1U9Q; -. DR PDBsum; 2AIM; -. DR PDBsum; 2OZ2; -. DR PDBsum; 3HD3; -. DR PDBsum; 3I06; -. DR PDBsum; 3IUT; -. DR PDBsum; 3KKU; -. DR PDBsum; 3LXS; -. DR PDBsum; 4KLB; -. DR PDBsum; 4PI3; -. DR PDBsum; 4QH6; -. DR PDBsum; 4W5B; -. DR PDBsum; 4W5C; -. DR PDBsum; 4XUI; -. DR PDBsum; 6N3S; -. DR PDBsum; 6O2X; -. DR PDBsum; 6UX6; -. DR PDBsum; 7JUJ; -. DR PDBsum; 7S18; -. DR PDBsum; 7S19; -. DR AlphaFoldDB; P25779; -. DR BMRB; P25779; -. DR SMR; P25779; -. DR BindingDB; P25779; -. DR ChEMBL; CHEMBL3563; -. DR DrugBank; DB02200; 3-[N-[benzyloxycarbonyl]-phenylalaninyl-amino]-5-phenyl-pentane-1-sulfonylmethylbenzene. DR DrugBank; DB04427; 4-Methyl-N-[(2S)-1-oxo-3-phenyl-1-[[(3S)-1-phenyl-5-(phenylmethoxysulfamoyl)pentan-3-yl]amino]propan-2-yl]piperazine-1-carboxamide. DR DrugBank; DB02051; 4-Nitrophenyl (3S)-3-({N-[(benzyloxy)carbonyl]-L-phenylalanyl}amino)-5-phenyl-1-pentanesulfonate. DR DrugBank; DB01871; [1-(1-Benzyl-3-Hydroxy-2-Oxo-Propylcarbamoyl)-2-Phenyl-Ethyl]-Carbamic Acid Benzyl Ester. DR DrugBank; DB01810; [1-(1-Methyl-4,5-Dioxo-Pent-2-Enylcarbamoyl)-2-Phenyl-Ethyl]-Carbamic Acid Benzyl Ester. DR DrugBank; DB02128; [1-(3-hydroxy-2-oxo-1-phenethyl-propylcarbamoyl)2-phenyl-ethyl]-carbamic acid pyridin-4-ylmethyl ester. DR DrugBank; DB03536; Benzyl N-[(2S)-5-(diaminomethylamino)-1-[[(2S)-4-fluoro-3-oxobutan-2-yl]amino]-1-oxopentan-2-yl]carbamate. DR DrugBank; DB03691; WRR-112. DR DrugBank; DB04502; WRR-204. DR DrugBank; DB03573; WRR-99. DR DrugCentral; P25779; -. DR MEROPS; C01.075; -. DR VEuPathDB; TriTrypDB:BCY84_22409; -. DR VEuPathDB; TriTrypDB:C3747_50g3; -. DR VEuPathDB; TriTrypDB:C3747_50g6; -. DR VEuPathDB; TriTrypDB:C3747_50g7; -. DR VEuPathDB; TriTrypDB:C4B63_89g68; -. DR VEuPathDB; TriTrypDB:ECC02_002401; -. DR VEuPathDB; TriTrypDB:Tc_MARK_8050; -. DR VEuPathDB; TriTrypDB:Tc_MARK_8695; -. DR VEuPathDB; TriTrypDB:TcBrA4_0068180; -. DR VEuPathDB; TriTrypDB:TcCL_NonESM02572; -. DR VEuPathDB; TriTrypDB:TcCLB.507603.260; -. DR VEuPathDB; TriTrypDB:TcCLB.509429.320; -. DR VEuPathDB; TriTrypDB:TCDM_05847; -. DR VEuPathDB; TriTrypDB:TcG_06121; -. DR VEuPathDB; TriTrypDB:TCSYLVIO_008967; -. DR VEuPathDB; TriTrypDB:TcYC6_0061420; -. DR VEuPathDB; TriTrypDB:TcYC6_0061430; -. DR VEuPathDB; TriTrypDB:TcYC6_0061460; -. DR VEuPathDB; TriTrypDB:TcYC6_0061470; -. DR BRENDA; 3.4.22.51; 6524. DR SABIO-RK; P25779; -. DR EvolutionaryTrace; P25779; -. DR GO; GO:0004197; F:cysteine-type endopeptidase activity; IEA:InterPro. DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW. DR CDD; cd02248; Peptidase_C1A; 1. DR Gene3D; 1.10.287.2250; -; 1. DR Gene3D; 3.90.70.10; Cysteine proteinases; 1. DR InterPro; IPR021981; DUF3586. DR InterPro; IPR038765; Papain-like_cys_pep_sf. DR InterPro; IPR025661; Pept_asp_AS. DR InterPro; IPR000169; Pept_cys_AS. DR InterPro; IPR025660; Pept_his_AS. DR InterPro; IPR013128; Peptidase_C1A. DR InterPro; IPR000668; Peptidase_C1A_C. DR InterPro; IPR039417; Peptidase_C1A_papain-like. DR InterPro; IPR013201; Prot_inhib_I29. DR PANTHER; PTHR12411:SF947; CATHEPSIN O; 1. DR PANTHER; PTHR12411; CYSTEINE PROTEASE FAMILY C1-RELATED; 1. DR Pfam; PF12131; DUF3586; 1. DR Pfam; PF08246; Inhibitor_I29; 1. DR Pfam; PF00112; Peptidase_C1; 1. DR PRINTS; PR00705; PAPAIN. DR SMART; SM00848; Inhibitor_I29; 1. DR SMART; SM00645; Pept_C1; 1. DR SUPFAM; SSF54001; Cysteine proteinases; 1. DR PROSITE; PS00640; THIOL_PROTEASE_ASN; 1. DR PROSITE; PS00139; THIOL_PROTEASE_CYS; 1. DR PROSITE; PS00639; THIOL_PROTEASE_HIS; 1. PE 1: Evidence at protein level; KW 3D-structure; Autocatalytic cleavage; Direct protein sequencing; KW Disulfide bond; Glycoprotein; Hydrolase; Protease; Signal; Thiol protease; KW Zymogen. FT SIGNAL 1..18 FT /evidence="ECO:0000305" FT PROPEP 19..122 FT /note="Activation peptide" FT /evidence="ECO:0000305|PubMed:2651912" FT /id="PRO_0000026372" FT CHAIN 123..467 FT /note="Cruzipain" FT /id="PRO_0000026373" FT REGION 333..355 FT /note="Disordered" FT /evidence="ECO:0000256|SAM:MobiDB-lite" FT ACT_SITE 147 FT ACT_SITE 284 FT ACT_SITE 304 FT SITE 337..338 FT /note="Cleavage; by autolysis" FT /evidence="ECO:0000269|PubMed:1559982" FT CARBOHYD 169 FT /note="N-linked (GlcNAc...) asparagine" FT /evidence="ECO:0000255" FT CARBOHYD 292 FT /note="N-linked (GlcNAc...) asparagine" FT /evidence="ECO:0000255" FT CARBOHYD 377 FT /note="N-linked (GlcNAc...) asparagine" FT /evidence="ECO:0000255" FT DISULFID 144..185 FT DISULFID 178..223 FT DISULFID 277..325 FT VARIANT 9 FT /note="L -> S (in clone 1800-2)" FT VARIANT 35 FT /note="T -> A (in clone 1800-2)" FT VARIANT 39 FT /note="A -> V (in clone 1800-2)" FT VARIANT 51 FT /note="S -> N (in clone 1800-4)" FT VARIANT 56 FT /note="A -> R (in clone 1800-2)" FT VARIANT 76 FT /note="N -> G (in clone 1800-4)" FT VARIANT 109 FT /note="Q -> G (in clone 1800-4)" FT VARIANT 117 FT /note="K -> N (in clone 1800-2)" FT VARIANT 146 FT /note="S -> G" FT VARIANT 157..158 FT /note="EC -> SG (in strain: RA)" FT VARIANT 186 FT /note="S -> G (in clones 1800-2 and 1800-4)" FT VARIANT 204 FT /note="A -> G (in strain: RA)" FT VARIANT 250..251 FT /note="WL -> CV (in clones 1800-2 and 1800-4)" FT VARIANT 261..262 FT /note="VD -> H (in strain: RA)" FT VARIANT 286 FT /note="V -> F (in clone 1800-4)" FT VARIANT 286 FT /note="V -> L (in strain: RA)" FT VARIANT 308 FT /note="T -> A (in clone 1800-2)" FT VARIANT 313 FT /note="E -> D" FT VARIANT 409 FT /note="L -> F (in clone 1800-2)" FT VARIANT 427 FT /note="K -> Q (in clone 1800-2)" FT VARIANT 430 FT /note="R -> W (in clone 1800-2)" FT VARIANT 457 FT /note="H -> Y (in clone 1800-2)" FT VARIANT 461 FT /note="H -> Q (in clone 1800-2)" FT CONFLICT 357 FT /note="S -> C (in Ref. 4)" FT /evidence="ECO:0000305" FT STRAND 125..128 FT /evidence="ECO:0007829|PDB:3I06" FT HELIX 129..132 FT /evidence="ECO:0007829|PDB:3I06" FT STRAND 143..145 FT /evidence="ECO:0007829|PDB:6UX6" FT HELIX 147..162 FT /evidence="ECO:0007829|PDB:3I06" FT HELIX 172..178 FT /evidence="ECO:0007829|PDB:3I06" FT STRAND 180..182 FT /evidence="ECO:0007829|PDB:3I06" FT HELIX 184..186 FT /evidence="ECO:0007829|PDB:3I06" FT HELIX 190..200 FT /evidence="ECO:0007829|PDB:3I06" FT STRAND 204..207 FT /evidence="ECO:0007829|PDB:3I06" FT TURN 208..210 FT /evidence="ECO:0007829|PDB:3I06" FT STRAND 216..218 FT /evidence="ECO:0007829|PDB:7S18" FT STRAND 225..227 FT /evidence="ECO:0007829|PDB:1F2A" FT STRAND 230..239 FT /evidence="ECO:0007829|PDB:3I06" FT HELIX 244..254 FT /evidence="ECO:0007829|PDB:3I06" FT STRAND 257..261 FT /evidence="ECO:0007829|PDB:3I06" FT HELIX 263..265 FT /evidence="ECO:0007829|PDB:4QH6" FT HELIX 266..268 FT /evidence="ECO:0007829|PDB:3I06" FT STRAND 271..274 FT /evidence="ECO:0007829|PDB:3I06" FT STRAND 284..297 FT /evidence="ECO:0007829|PDB:3I06" FT STRAND 299..303 FT /evidence="ECO:0007829|PDB:3I06" FT TURN 307..309 FT /evidence="ECO:0007829|PDB:7S18" FT STRAND 315..322 FT /evidence="ECO:0007829|PDB:3I06" FT HELIX 324..326 FT /evidence="ECO:0007829|PDB:3I06" FT STRAND 329..336 FT /evidence="ECO:0007829|PDB:3I06" SQ SEQUENCE 467 AA; 49836 MW; B93EBA49B511363D CRC64; MSGWARALLL AAVLVVMACL VPAATASLHA EETLTSQFAE FKQKHGRVYE SAAEEAFRLS VFRENLFLAR LHAAANPHAT FGVTPFSDLT REEFRSRYHN GAAHFAAAQE RARVPVKVEV VGAPAAVDWR ARGAVTAVKD QGQCGSCWAF SAIGNVECQW FLAGHPLTNL SEQMLVSCDK TDSGCSGGLM NNAFEWIVQE NNGAVYTEDS YPYASGEGIS PPCTTSGHTV GATITGHVEL PQDEAQIAAW LAVNGPVAVA VDASSWMTYT GGVMTSCVSE QLDHGVLLVG YNDSAAVPYW IIKNSWTTQW GEEGYIRIAK GSNQCLVKEE ASSAVVGGPG PTPEPTTTTT TSAPGPSPSY FVQMSCTDAA CIVGCENVTL PTGQCLLTTS GVSAIVTCGA ETLTEEVFLT STHCSGPSVR SSVPLNKCNR LLRGSVEFFC GSSSSGRLAD VDRQRRHQPY HSRHRRL //