P46541 (PIP_BACCO) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Proline iminopeptidase
Short name=PIP
EC=3.4.11.5

Alternative name(s):
Prolyl aminopeptidase
Short name=PAP

Gene names

Name:

pip

Organism

Bacillus coagulans

Taxonomic identifier

1398 [NCBI]

Taxonomic lineage

Bacteria › Firmicutes › Bacillales › Bacillaceae › Bacillus

Protein attributes

Sequence length	288 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Releases the N-terminal proline from various substrates including at least dipeptides Pro-Pro, Pro-Gln, Pro-Trp and Pro-Tyr. Acts also on amides (Pro-beta NA) and oligopeptides including Pro-Leu-GlyNH2, Pro-Leu-Gly, Pro-Phe-Gly-Lys, Pro-Pro-Ala-OBut and Pro-Pro-Gly-(Pro-Pro-Gly)4. Higher activity toward small peptides (up to three residues), but very low activity for longer peptides. Has no activity against p-nitrophenyl acetate, poly_L-proline, Met-Pro or amino acyl amides other than Pro-betaNA (Pyr-betaNA, Phe-betaNA, Cys-betaNA, Met-betaNA, Leu-betaNA, Ala-betaNA and Z-Gly-Pro-betaNA). Ref.1 Ref.3
Catalytic activity	Release of N-terminal proline from a peptide. Ref.1 Ref.3 Ref.4
Enzyme regulation	Completely inhibited by p-chloromercuribenzoate (PCMB) and heavy metal salts. Partially inhibited by proline and proline derivatives with proline as the amino terminus. Enzyme inactivated by PCMB is reactivated by incubation with 2-mercaptoethanol. Ref.1 Ref.3
Subunit structure	Monomer. Ref.3
Subcellular location	Cytoplasm Probable.
Sequence similarities	Belongs to the peptidase S33 family.
Biophysicochemical properties	Kinetic parameters: KM=4.08 mM for Pro-Ala Ref.1 Ref.3 KM=1.16 mM for Pro-Asp KM=1.06 mM for Pro-Gly KM=0.91 mM for Pro-Lys KM=7.70 mM for Pro-Leu KM=1.50 mM for Pro-Phe KM=1.27 mM for Pro-Trp KM=0.95 mM for Pro-Tyr KM=0.26 mM for Pro-2-NNap KM=0.26 mM for Pro-pNA KM=27.0 mM for Pro-D-Ala KM=2.51 mM for Pro-D-Phe KM=6.35 mM for Pro-Leu-Gly-NH2 pH dependence: Optimum pH is 8.0. Stable at pH 5.5-7.5. Most active at 7.3 with Pro-beta-naphthylamide (Pro-2-NNap) as the substrate. More than 85% of the activity remains between pH 6.5 and 7.5 after incubation for 15 minutes at 30 degrees Celsius and it retains 50% of the original activity after incubation at pH 7.0 and at 30 degrees Celsius for 30 minutes. 50% of the activity remains after 15 minutes preincubations at pH 8.0. Temperature dependence: Maximum activity at 40 degrees Celsius. Stable at temperatures up to 38 degrees Celsius.

Ontologies

Keywords
Cellular component	Cytoplasm
Molecular function	Aminopeptidase Hydrolase Protease
Technical term	Direct protein sequencing
Gene Ontology (GO)
Biological process	proteolysis Inferred from electronic annotation. Source: UniProtKB-KW
Cellular component	cytoplasm Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular function	aminopeptidase activity Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Chain	1 – 288	288	Proline iminopeptidase		PRO_0000080835
Sites
	Active site	101	1	Nucleophile Probable
	Active site	240	1	By similarity
	Active site	267	1	Proton donor By similarity
Experimental info
	Mutagenesis	62	1	C → S: Activity same as wild-type. Ref.4
	Mutagenesis	101	1	S → A: Complete inactivation. Ref.4

Sequences

	Sequence		Length	Mass (Da)
	P46541 [UniParc]. Last modified October 1, 1996. Version 2. Checksum: 80AD8D594CB648C6	FASTA	288	32,357
	10 20 30 40 50 60 MYTEGFIDVT GGRVSFQKFD ENGGGTPVIV LHGGPGSSCY SLLGLKALAK DRPVILYDQL 70 80 90 100 110 120 GCGKSDRPMD TTLWRLDRFV EELAQIRQAL NLDEVHILGH SWGTTLAAAY CLTKPSGVKS 130 140 150 160 170 180 VIFSSPCLSA PLWEQDQKRN LKKLPLDVQE TINRCEENGT TDSEEFAAAI EVFGKHFVNR 190 200 210 220 230 240 LEKQPEWLEQ KPSGYRNADI YNIMWGPSEF TVLGNLKNFD CTTQLKEITC PSLYTCGRFD 250 260 270 280 EATPETTEYY SSLTPKSKFH VFEKSAHMPY IEEPEEYLAV IGDFLNSI

References

[1]	"Cloning, sequencing, and high expression of the proline iminopeptidase gene from Bacillus coagulans." Kitazono A., Yoshimoto T., Tsuru D. J. Bacteriol. 174:7919-7925(1992) [PubMed: 1459939] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-13; 20-23; 51-56; 66-73; 93-95; 182-184; 199-202 AND 283-288, CRYSTALLIZATION, FUNCTION, CATALYTIC ACTIVITY, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES.
[2]	Yoshimoto T. Submitted (APR-1994) to the EMBL/GenBank/DDBJ databases Cited for: SEQUENCE REVISION.
[3]	"Proline iminopeptidase from Bacillus coagulans: purification and enzymatic properties." Yoshimoto T., Tsuru D. J. Biochem. 97:1477-1485(1985) [PubMed: 4030733] [Abstract] Cited for: FUNCTION, CATALYTIC ACTIVITY, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT.
[4]	"Prolyl aminopeptidase is not a sulfhydryl enzyme: identification of the active serine residue by site-directed mutagenesis." Kitazono A., Ito K., Yoshimoto T. J. Biochem. 116:943-945(1994) [PubMed: 7896753] [Abstract] Cited for: CATALYTIC ACTIVITY, MUTAGENESIS OF CYS-62 AND SER-101.
+	Additional computationally mapped references.

Cross-references

Sequence databases
EMBL GenBank DDBJ	D11037 Genomic DNA. Translation: BAA01792.1.
3D structure databases
ProteinModelPortal	P46541.
ModBase	Search...
Protein family/group databases
MEROPS	S33.010.
Protocols and materials databases
StructuralBiologyKnowledgebase	Search...
Family and domain databases
InterPro	IPR000073. AB_hydrolase_1. IPR005945. Pept_S33_TRI_F1. IPR002410. Peptidase_S33. [Graphical view]
Pfam	PF00561. Abhydrolase_1. 1 hit. [Graphical view]
PIRSF	PIRSF005539. Pept_S33_TRI_F1. 1 hit.
PRINTS	PR00793. PROAMNOPTASE.
TIGRFAMs	TIGR01250. Pro_imino_pep_2. 1 hit.
ProtoNet	Search...

Entry information

Entry name

PIP_BACCO

Accession

Primary (citable) accession number: P46541

Entry history

Integrated into UniProtKB/Swiss-Prot:	November 1, 1995
Last sequence update:	October 1, 1996
Last modified:	November 16, 2011
This is version 66 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program