ID PAPA4_CARPA Reviewed; 348 AA. AC P05994; DT 01-NOV-1988, integrated into UniProtKB/Swiss-Prot. DT 01-OCT-1996, sequence version 3. DT 27-MAR-2024, entry version 125. DE RecName: Full=Papaya proteinase 4; DE EC=3.4.22.25 {ECO:0000269|PubMed:2404797}; DE AltName: Full=Glycyl endopeptidase; DE AltName: Full=Papaya peptidase B; DE AltName: Full=Papaya proteinase IV; DE Short=PPIV; DE Flags: Precursor; OS Carica papaya (Papaya). OC Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; OC Spermatophyta; Magnoliopsida; eudicotyledons; Gunneridae; Pentapetalae; OC rosids; malvids; Brassicales; Caricaceae; Carica. OX NCBI_TaxID=3649; RN [1] RP NUCLEOTIDE SEQUENCE [MRNA]. RC TISSUE=Leaf; RX PubMed=7770454; DOI=10.1093/protein/8.1.59; RA Taylor M.A., Baker K.C., Briggs G.S., Connerton I.F., Cummings N.J., RA Pratt K.A., Revell D.F., Freedman R.B., Goodenough P.W.; RT "Recombinant pro-regions from papain and papaya proteinase IV-are selective RT high affinity inhibitors of the mature papaya enzymes."; RL Protein Eng. 8:59-62(1995). RN [2] RP PROTEIN SEQUENCE OF 133-348. RX PubMed=2591528; DOI=10.1016/0014-5793(89)81627-8; RA Ritonja A., Buttle D.J., Rawlings N.D., Turk V., Barrett A.J.; RT "Papaya proteinase IV amino acid sequence."; RL FEBS Lett. 258:109-112(1989). RN [3] RP PROTEIN SEQUENCE OF 133-150. RX PubMed=518921; DOI=10.1016/0005-2795(79)90257-5; RA Lynn K.R., Yaguchi M.; RT "N-terminal homology in three cysteinyl proteases from Papaya latex."; RL Biochim. Biophys. Acta 581:363-364(1979). RN [4] RP NUCLEOTIDE SEQUENCE [MRNA] OF 287-348. RX PubMed=3541893; DOI=10.1042/bj2370105; RA McKee R.A., Adams S., Matthews J.A., Smith C.J., Smith H.; RT "Molecular cloning of two cysteine proteinases from paw-paw (Carica RT papaya)."; RL Biochem. J. 237:105-110(1986). RN [5] RP FUNCTION, SUBSTRATE SPECIFICITY, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL RP PROPERTIES. RX PubMed=2404797; DOI=10.1016/0014-5793(90)80101-n; RA Buttle D.J., Ritonja A., Pearl L.H., Turk V., Barrett A.J.; RT "Selective cleavage of glycyl bonds by papaya proteinase IV."; RL FEBS Lett. 260:195-197(1990). RN [6] RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF 133-348, AND DISULFIDE BONDS. RX PubMed=7548082; DOI=10.1021/bi00040a034; RA O'Hara B.P., Hemmings A.M., Buttle D.J., Pearl L.H.; RT "Crystal structure of glycyl endopeptidase from Carica papaya: a cysteine RT endopeptidase of unusual substrate specificity."; RL Biochemistry 34:13190-13195(1995). CC -!- FUNCTION: Thiol protease with a substrate specificity very different CC from the other thiol proteases. {ECO:0000269|PubMed:2404797}. CC -!- CATALYTIC ACTIVITY: CC Reaction=Preferential cleavage: Gly-|-Xaa, in proteins and in small CC molecule substrates.; EC=3.4.22.25; CC Evidence={ECO:0000269|PubMed:2404797}; CC -!- ACTIVITY REGULATION: Not inhibited by cystatin. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=5.2 mM for Boc-Ala-Ala-Gly-NHPhNO(2) {ECO:0000269|PubMed:2404797}; CC KM=0.16 mM for Boc-Ala-Ala-Gly-NHMec {ECO:0000269|PubMed:2404797}; CC KM=0.08 mM for Boc-Ala-Ala-Ala-NHMec {ECO:0000269|PubMed:2404797}; CC Note=kcat is 22 sec(-1) with Boc-Ala-Ala-Gly-NHPhNO(2) as substrate CC (PubMed:2404797). kcat is 5 sec(-1) with Boc-Ala-Ala-Gly-NHMec as CC substrate (PubMed:2404797). kcat is 0.08 sec(-1) with CC Boc-Ala-Ala-Ala-NHMec as substrate (PubMed:2404797). CC {ECO:0000269|PubMed:2404797}; CC -!- MISCELLANEOUS: Substitution of the conserved Gly residue by Glu-155 CC could possibly explain the unusual specificity. 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; X78056; CAA54974.1; -; mRNA. DR EMBL; X03970; CAA27608.1; -; mRNA. DR PIR; S06837; S06837. DR PIR; T09798; T09798. DR PDB; 1GEC; X-ray; 2.10 A; E=133-348. DR PDBsum; 1GEC; -. DR AlphaFoldDB; P05994; -. DR SMR; P05994; -. DR Allergome; 1539; Cari p Endoproteinase. DR MEROPS; C01.004; -. DR MEROPS; I29.003; -. DR KEGG; ag:CAA54974; -. DR BRENDA; 3.4.22.25; 1191. DR SABIO-RK; P05994; -. DR EvolutionaryTrace; P05994; -. DR GO; GO:0008234; F:cysteine-type peptidase activity; IEA:UniProtKB-KW. DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW. DR CDD; cd02248; Peptidase_C1A; 1. DR Gene3D; 3.90.70.10; Cysteine proteinases; 1. 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; CYSTEINE PROTEASE FAMILY C1-RELATED; 1. DR PANTHER; PTHR12411:SF357; OS01G0971400 PROTEIN; 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; Direct protein sequencing; Disulfide bond; Hydrolase; KW Protease; Signal; Thiol protease; Zymogen. FT SIGNAL 1..18 FT /evidence="ECO:0000255" FT PROPEP 19..132 FT /note="Activation peptide" FT /evidence="ECO:0000269|PubMed:2591528, FT ECO:0000269|PubMed:518921" FT /id="PRO_0000026412" FT CHAIN 133..348 FT /note="Papaya proteinase 4" FT /id="PRO_0000026413" FT ACT_SITE 157 FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10088" FT ACT_SITE 291 FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10089" FT ACT_SITE 311 FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10090" FT DISULFID 154..195 FT /evidence="ECO:0000269|PubMed:7548082, FT ECO:0007744|PDB:1GEC" FT DISULFID 188..227 FT /evidence="ECO:0000269|PubMed:7548082, FT ECO:0007744|PDB:1GEC" FT DISULFID 285..336 FT /evidence="ECO:0000269|PubMed:7548082, FT ECO:0007744|PDB:1GEC" FT CONFLICT 190 FT /note="K -> L (in Ref. 2; AA sequence)" FT /evidence="ECO:0000305" FT TURN 139..143 FT /evidence="ECO:0007829|PDB:1GEC" FT STRAND 153..155 FT /evidence="ECO:0007829|PDB:1GEC" FT HELIX 157..174 FT /evidence="ECO:0007829|PDB:1GEC" FT HELIX 182..188 FT /evidence="ECO:0007829|PDB:1GEC" FT STRAND 190..192 FT /evidence="ECO:0007829|PDB:1GEC" FT HELIX 200..210 FT /evidence="ECO:0007829|PDB:1GEC" FT TURN 215..217 FT /evidence="ECO:0007829|PDB:1GEC" FT HELIX 229..232 FT /evidence="ECO:0007829|PDB:1GEC" FT STRAND 241..244 FT /evidence="ECO:0007829|PDB:1GEC" FT HELIX 250..257 FT /evidence="ECO:0007829|PDB:1GEC" FT STRAND 262..266 FT /evidence="ECO:0007829|PDB:1GEC" FT HELIX 271..274 FT /evidence="ECO:0007829|PDB:1GEC" FT STRAND 278..281 FT /evidence="ECO:0007829|PDB:1GEC" FT STRAND 291..301 FT /evidence="ECO:0007829|PDB:1GEC" FT STRAND 304..310 FT /evidence="ECO:0007829|PDB:1GEC" FT STRAND 322..326 FT /evidence="ECO:0007829|PDB:1GEC" FT HELIX 335..337 FT /evidence="ECO:0007829|PDB:1GEC" FT STRAND 343..346 FT /evidence="ECO:0007829|PDB:1GEC" SQ SEQUENCE 348 AA; 39024 MW; 40855EDD37F68A8E CRC64; MAIICSFSKL LFVAICLFGH MSLSYCDFSI VGYSQDDLTS TERLIQLFNS WMLKHNKNYK NVDEKLYRFE IFKDNLKYID ERNKMINGYW LGLNEFSDLS NDEFKEKYVG SLPEDYTNQP YDEEFVNEDI VDLPESVDWR AKGAVTPVKH QGYCESCWAF STVATVEGIN KIKTGNLVEL SEQELVDCDK QSYGCNRGYQ STSLQYVAQN GIHLRAKYPY IAKQQTCRAN QVGGPKVKTN GVGRVQSNNE GSLLNAIAHQ PVSVVVESAG RDFQNYKGGI FEGSCGTKVD HAVTAVGYGK SGGKGYILIK NSWGPGWGEN GYIRIRRASG NSPGVCGVYR SSYYPIKN //