PIP_LACDE
ID PIP_LACDE Reviewed; 295 AA.
AC P46544;
DT 01-NOV-1995, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-1996, sequence version 2.
DT 03-AUG-2022, entry version 105.
DE RecName: Full=Proline iminopeptidase;
DE Short=PIP;
DE EC=3.4.11.5;
DE AltName: Full=Prolyl aminopeptidase;
DE Short=PAP;
GN Name=pepIP;
OS Lactobacillus delbrueckii subsp. bulgaricus.
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Lactobacillaceae;
OC Lactobacillus.
OX NCBI_TaxID=1585;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-11, CATALYTIC
RP ACTIVITY, AND SUBCELLULAR LOCATION.
RC STRAIN=CNRZ 397;
RX PubMed=8012575; DOI=10.1099/00221287-140-3-527;
RA Atlan D., Gilbert C., Blanc B., Portalier R.;
RT "Cloning, sequencing and characterization of the pepIP gene encoding a
RT proline iminopeptidase from Lactobacillus delbrueckii subsp. bulgaricus
RT CNRZ 397.";
RL Microbiology 140:527-535(1994).
RN [2]
RP PROTEIN SEQUENCE OF 1-11, FUNCTION, CATALYTIC ACTIVITY, ACTIVITY
RP REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY, AND
RP SUBUNIT.
RC STRAIN=CNRZ 397;
RX PubMed=8012576; DOI=10.1099/00221287-140-3-537;
RA Gilbert C., Atlan D., Blanc B., Portalier R.;
RT "Proline iminopeptidase from Lactobacillus delbrueckii subsp. bulgaricus
RT CNRZ 397: purification and characterization.";
RL Microbiology 140:537-542(1994).
RN [3]
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, CIRCULAR DICHROISM, AND
RP MUTAGENESIS OF HIS-34; GLY-35; GLY-36; PRO-37; GLY-38; ASP-62; GLY-65;
RP SER-69; PRO-72; GLU-88; GLY-105; GLN-106; SER-107; TRP-108; GLY-109;
RP GLY-110; SER-132; GLU-143; PRO-237; GLY-243; ASP-245; ASP-246 AND HIS-273.
RX PubMed=9989236; DOI=10.1016/s0167-4838(98)00264-7;
RA Morel F., Gilbert C., Geourjon C., Frot-Coutaz J., Portalier R., Atlan D.;
RT "The prolyl aminopeptidase from Lactobacillus delbrueckii subsp. bulgaricus
RT belongs to the alpha/beta hydrolase fold family.";
RL Biochim. Biophys. Acta 1429:501-505(1999).
CC -!- FUNCTION: Releases the N-terminal proline from various substrates. Has
CC a high specificity towards di- or tripeptides with proline at the
CC NH(2)-terminal position, but is not able to hydrolyze longer peptides,
CC or peptides with hydroxyproline at the NH(2)-end. Partially hydrolyzes
CC also peptides with alanine, glycine and leucine at the NH(2)-terminal
CC position. {ECO:0000269|PubMed:8012576}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Release of N-terminal proline from a peptide.; EC=3.4.11.5;
CC Evidence={ECO:0000269|PubMed:8012575, ECO:0000269|PubMed:8012576,
CC ECO:0000269|PubMed:9989236};
CC -!- ACTIVITY REGULATION: Inhibited strongly by 3,4-dichloroisocoumarin,
CC bestatin and heavy metal ions. Inactivated by p-chloromercuribenzoate,
CC but reactivated by dithiothreitol. {ECO:0000269|PubMed:8012576}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.93 mM for prolyl-pNA (at 30 degrees Celsius and pH 7.0)
CC {ECO:0000269|PubMed:8012576, ECO:0000269|PubMed:9989236};
CC Vmax=738 mmol/min/mg enzyme with prolyl-pNA as substrate (at 30
CC degrees Celsius and pH 7.0) {ECO:0000269|PubMed:8012576,
CC ECO:0000269|PubMed:9989236};
CC pH dependence:
CC Optimum pH is 6-7. {ECO:0000269|PubMed:8012576,
CC ECO:0000269|PubMed:9989236};
CC Temperature dependence:
CC Stable at temperatures below 40 degrees Celsius.
CC {ECO:0000269|PubMed:8012576, ECO:0000269|PubMed:9989236};
CC -!- SUBUNIT: Homotrimer. {ECO:0000269|PubMed:8012576}.
CC -!- SUBCELLULAR LOCATION: Cell envelope {ECO:0000269|PubMed:8012575}.
CC -!- SIMILARITY: Belongs to the peptidase S33 family. {ECO:0000305}.
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DR EMBL; L10712; AAA61596.1; -; Genomic_DNA.
DR AlphaFoldDB; P46544; -.
DR SMR; P46544; -.
DR ESTHER; lacdl-pip; Proline_iminopeptidase.
DR MEROPS; S33.021; -.
DR SABIO-RK; P46544; -.
DR GO; GO:0031975; C:envelope; 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; Direct protein sequencing; Hydrolase; Protease.
FT CHAIN 1..295
FT /note="Proline iminopeptidase"
FT /id="PRO_0000080836"
FT DOMAIN 29..279
FT /note="AB hydrolase-1"
FT /evidence="ECO:0000255"
FT ACT_SITE 107
FT /note="Nucleophile"
FT /evidence="ECO:0000250"
FT ACT_SITE 246
FT /evidence="ECO:0000250"
FT ACT_SITE 273
FT /note="Proton donor"
FT /evidence="ECO:0000250"
FT MUTAGEN 34
FT /note="H->Q: 0.1% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 35
FT /note="G->D: 0.1% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 36
FT /note="G->E: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 37
FT /note="P->L: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 38
FT /note="G->D: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 62
FT /note="D->G: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 65
FT /note="G->D: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 69
FT /note="S->F: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 72
FT /note="P->A: 95.4% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 88
FT /note="E->K: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 105
FT /note="G->E: 0.2% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 106
FT /note="Q->L: 21.8% of wild-type activity. KM=0.2 mM for
FT prolyl-pNA."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 107
FT /note="S->G,N: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 108
FT /note="W->Y: 37.9% of wild-type activity. KM=4.0 mM for
FT prolyl-pNA."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 109
FT /note="G->A: 0.2% of wild-type activity. KM=2.3 mM for
FT prolyl-pNA."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 109
FT /note="G->S,D: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 110
FT /note="G->E: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 132
FT /note="S->F: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 143
FT /note="E->K: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 237
FT /note="P->A: 89.2% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 243
FT /note="G->D: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 245
FT /note="D->G: 22.5% of wild-type activity. KM=1.0 mM for
FT prolyl-pNA."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 246
FT /note="D->G: 0.1% of wild-type activity. KM=1.0 mM for
FT prolyl-pNA."
FT /evidence="ECO:0000269|PubMed:9989236"
FT MUTAGEN 273
FT /note="H->Q: Loss of activity."
FT /evidence="ECO:0000269|PubMed:9989236"
SQ SEQUENCE 295 AA; 33027 MW; 97CF3AAD9FAFF900 CRC64;
MMQITEKYLP FGNWQTYCRI VGEATDRAPL LLLHGGPGSS HNYFEVLDQV AEKSGRQVIM
YDQLGCGNSS IPDDQAETAY TAQTWVKELE NVREQLGLDQ IHLLGQSWGG MLALIYLCDY
QPEGVKSLIL SSTLASAKLW SQELHRLIKY LPKGEQAAIK EAETTGNYDS LAYQAANAHF
MDQHAIKLTP DLPEPVLRKK KGGSLAYLTG WGPNEYTPIG NLHGYEYTDR LKDLHLPALI
TSGTDDLCTP LVAKSMYDNL PNARWELFAG CGHMPFVQEN AKYQELLSDW LISQD
