PIP_WEICA
ID PIP_WEICA Reviewed; 288 AA.
AC P46541;
DT 01-NOV-1995, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-1996, sequence version 2.
DT 03-AUG-2022, entry version 91.
DE RecName: Full=Proline iminopeptidase;
DE Short=PIP;
DE EC=3.4.11.5;
DE AltName: Full=Prolyl aminopeptidase;
DE Short=PAP;
GN Name=pip;
OS Weizmannia coagulans (Bacillus coagulans).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Weizmannia.
OX NCBI_TaxID=1398;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-13; 20-23; 51-56;
RP 66-73; 93-95; 182-184; 199-202 AND 283-288, CRYSTALLIZATION, FUNCTION,
RP CATALYTIC ACTIVITY, ACTIVITY REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=1459939; DOI=10.1128/jb.174.24.7919-7925.1992;
RA Kitazono A., Yoshimoto T., Tsuru D.;
RT "Cloning, sequencing, and high expression of the proline iminopeptidase
RT gene from Bacillus coagulans.";
RL J. Bacteriol. 174:7919-7925(1992).
RN [2]
RP SEQUENCE REVISION.
RA Yoshimoto T.;
RL Submitted (APR-1994) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND SUBUNIT.
RX PubMed=4030733; DOI=10.1093/oxfordjournals.jbchem.a135202;
RA Yoshimoto T., Tsuru D.;
RT "Proline iminopeptidase from Bacillus coagulans: purification and enzymatic
RT properties.";
RL J. Biochem. 97:1477-1485(1985).
RN [4]
RP CATALYTIC ACTIVITY, AND MUTAGENESIS OF CYS-62 AND SER-101.
RX PubMed=7896753; DOI=10.1093/oxfordjournals.jbchem.a124649;
RA Kitazono A., Ito K., Yoshimoto T.;
RT "Prolyl aminopeptidase is not a sulfhydryl enzyme: identification of the
RT active serine residue by site-directed mutagenesis.";
RL J. Biochem. 116:943-945(1994).
CC -!- FUNCTION: Releases the N-terminal proline from various substrates
CC including at least dipeptides Pro-Pro, Pro-Gln, Pro-Trp and Pro-Tyr.
CC Acts also on amides (Pro-beta NA) and oligopeptides including Pro-Leu-
CC GlyNH2, Pro-Leu-Gly, Pro-Phe-Gly-Lys, Pro-Pro-Ala-OBut and Pro-Pro-Gly-
CC (Pro-Pro-Gly)(4). Higher activity toward small peptides (up to three
CC residues), but very low activity for longer peptides. Has no activity
CC against p-nitrophenyl acetate, poly_L-proline, Met-Pro or amino acyl
CC amides other than Pro-betaNA (Pyr-betaNA, Phe-betaNA, Cys-betaNA, Met-
CC betaNA, Leu-betaNA, Ala-betaNA and Z-Gly-Pro-betaNA).
CC {ECO:0000269|PubMed:1459939, ECO:0000269|PubMed:4030733}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Release of N-terminal proline from a peptide.; EC=3.4.11.5;
CC Evidence={ECO:0000269|PubMed:1459939, ECO:0000269|PubMed:4030733,
CC ECO:0000269|PubMed:7896753};
CC -!- ACTIVITY REGULATION: Completely inhibited by p-chloromercuribenzoate
CC (PCMB) and heavy metal salts. Partially inhibited by proline and
CC proline derivatives with proline as the amino terminus. Enzyme
CC inactivated by PCMB is reactivated by incubation with 2-
CC mercaptoethanol. {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=4.08 mM for Pro-Ala {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=1.16 mM for Pro-Asp {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=1.06 mM for Pro-Gly {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=0.91 mM for Pro-Lys {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=7.70 mM for Pro-Leu {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=1.50 mM for Pro-Phe {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=1.27 mM for Pro-Trp {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=0.95 mM for Pro-Tyr {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=0.26 mM for Pro-2-NNap {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=0.26 mM for Pro-pNA {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=27.0 mM for Pro-D-Ala {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=2.51 mM for Pro-D-Phe {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC KM=6.35 mM for Pro-Leu-Gly-NH(2) {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC pH dependence:
CC Optimum pH is 8.0. Stable at pH 5.5-7.5. Most active at 7.3 with Pro-
CC beta-naphthylamide (Pro-2-NNap) as the substrate. More than 85% of
CC the activity remains between pH 6.5 and 7.5 after incubation for 15
CC minutes at 30 degrees Celsius and it retains 50% of the original
CC activity after incubation at pH 7.0 and at 30 degrees Celsius for 30
CC minutes. 50% of the activity remains after 15 minutes preincubations
CC at pH 8.0. {ECO:0000269|PubMed:1459939, ECO:0000269|PubMed:4030733};
CC Temperature dependence:
CC Maximum activity at 40 degrees Celsius. Stable at temperatures up to
CC 38 degrees Celsius. {ECO:0000269|PubMed:1459939,
CC ECO:0000269|PubMed:4030733};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:4030733}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the peptidase S33 family. {ECO:0000305}.
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DR EMBL; D11037; BAA01792.1; -; Genomic_DNA.
DR AlphaFoldDB; P46541; -.
DR SMR; P46541; -.
DR ChEMBL; CHEMBL2390809; -.
DR ESTHER; bacco-pip; Proline_iminopeptidase.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0004177; F:aminopeptidase activity; IEA:UniProtKB-KW.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR000073; AB_hydrolase_1.
DR InterPro; IPR002410; Peptidase_S33.
DR InterPro; IPR005945; Pro_imino_pep.
DR Pfam; PF00561; Abhydrolase_1; 1.
DR PIRSF; PIRSF005539; Pept_S33_TRI_F1; 1.
DR PRINTS; PR00793; PROAMNOPTASE.
DR SUPFAM; SSF53474; SSF53474; 1.
DR TIGRFAMs; TIGR01250; pro_imino_pep_2; 1.
PE 1: Evidence at protein level;
KW Aminopeptidase; Cytoplasm; Direct protein sequencing; Hydrolase; Protease.
FT CHAIN 1..288
FT /note="Proline iminopeptidase"
FT /id="PRO_0000080835"
FT DOMAIN 27..274
FT /note="AB hydrolase-1"
FT /evidence="ECO:0000255"
FT ACT_SITE 101
FT /note="Nucleophile"
FT /evidence="ECO:0000305"
FT ACT_SITE 240
FT /evidence="ECO:0000250"
FT ACT_SITE 267
FT /note="Proton donor"
FT /evidence="ECO:0000250"
FT MUTAGEN 62
FT /note="C->S: Activity same as wild-type."
FT /evidence="ECO:0000269|PubMed:7896753"
FT MUTAGEN 101
FT /note="S->A: Complete inactivation."
FT /evidence="ECO:0000269|PubMed:7896753"
SQ SEQUENCE 288 AA; 32357 MW; 80AD8D594CB648C6 CRC64;
MYTEGFIDVT GGRVSFQKFD ENGGGTPVIV LHGGPGSSCY SLLGLKALAK DRPVILYDQL
GCGKSDRPMD TTLWRLDRFV EELAQIRQAL NLDEVHILGH SWGTTLAAAY CLTKPSGVKS
VIFSSPCLSA PLWEQDQKRN LKKLPLDVQE TINRCEENGT TDSEEFAAAI EVFGKHFVNR
LEKQPEWLEQ KPSGYRNADI YNIMWGPSEF TVLGNLKNFD CTTQLKEITC PSLYTCGRFD
EATPETTEYY SSLTPKSKFH VFEKSAHMPY IEEPEEYLAV IGDFLNSI
