APX_STRGG
ID APX_STRGG Reviewed; 445 AA.
AC P80561; B1W291;
DT 01-FEB-1996, integrated into UniProtKB/Swiss-Prot.
DT 26-NOV-2014, sequence version 2.
DT 03-AUG-2022, entry version 122.
DE RecName: Full=Aminopeptidase S;
DE EC=3.4.11.24;
DE AltName: Full=API {ECO:0000303|PubMed:2503378};
DE AltName: Full=SGAP {ECO:0000303|PubMed:9048953};
DE Flags: Precursor;
GN OrderedLocusNames=SGR_5809;
OS Streptomyces griseus subsp. griseus (strain JCM 4626 / NBRC 13350).
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=455632;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=JCM 4626 / NBRC 13350;
RX PubMed=18375553; DOI=10.1128/jb.00204-08;
RA Ohnishi Y., Ishikawa J., Hara H., Suzuki H., Ikenoya M., Ikeda H.,
RA Yamashita A., Hattori M., Horinouchi S.;
RT "Genome sequence of the streptomycin-producing microorganism Streptomyces
RT griseus IFO 13350.";
RL J. Bacteriol. 190:4050-4060(2008).
RN [2]
RP PROTEIN SEQUENCE OF 46-329, FUNCTION, AND MASS SPECTROMETRY.
RX PubMed=8665903; DOI=10.1111/j.1432-1033.1996.00843.x;
RA Maras B., Greenblatt H.M., Shoham G., Spungin-Bialik A., Blumberg S.,
RA Barra D.;
RT "Aminopeptidase from Streptomyces griseus: primary structure and comparison
RT with other zinc-containing aminopeptidases.";
RL Eur. J. Biochem. 236:843-846(1996).
RN [3]
RP PROTEIN SEQUENCE OF 46-51, FUNCTION, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, SUBCELLULAR LOCATION, AND THERMAL
RP STABILITY.
RX PubMed=2503378; DOI=10.1111/j.1432-1033.1989.tb14952.x;
RA Spungin A., Blumberg S.;
RT "Streptomyces griseus aminopeptidase is a calcium-activated zinc
RT metalloprotein. Purification and properties of the enzyme.";
RL Eur. J. Biochem. 183:471-477(1989).
RN [4]
RP CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, COFACTOR, ACTIVITY REGULATION,
RP AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=8444149; DOI=10.1111/j.1432-1033.1993.tb17639.x;
RA Ben-Meir D., Spungin A., Ashkenazi R., Blumberg S.;
RT "Specificity of Streptomyces griseus aminopeptidase and modulation of
RT activity by divalent metal ion binding and substitution.";
RL Eur. J. Biochem. 212:107-112(1993).
RN [5]
RP SEQUENCE REVISION TO 115 AND 229, FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES,
RP POSSIBLE REACTION MECHANISM, ACTIVE SITE, MUTAGENESIS OF GLU-176 AND
RP TYR-291, AND EXPRESSION IN E.COLI.
RC STRAIN=DSM 40855;
RX PubMed=15280041; DOI=10.1016/j.febslet.2004.07.001;
RA Fundoiano-Hershcovitz Y., Rabinovitch L., Langut Y., Reiland V., Shoham G.,
RA Shoham Y.;
RT "Identification of the catalytic residues in the double-zinc aminopeptidase
RT from Streptomyces griseus.";
RL FEBS Lett. 571:192-196(2004).
RN [6]
RP FUNCTION, ACTIVITY REGULATION, REACTION MECHANISM, AND ACTIVE SITE.
RX PubMed=17608735; DOI=10.1111/j.1742-4658.2007.05912.x;
RA Hershcovitz Y.F., Gilboa R., Reiland V., Shoham G., Shoham Y.;
RT "Catalytic mechanism of SGAP, a double-zinc aminopeptidase from
RT Streptomyces griseus.";
RL FEBS J. 274:3864-3876(2007).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS) IN COMPLEX WITH CALCIUM AND ZINC,
RP COFACTOR, SUBUNIT, AND DISULFIDE BOND.
RX PubMed=9048953; DOI=10.1006/jmbi.1996.0729;
RA Greenblatt H.M., Almog O., Maras B., Spungin-Bialik A., Barra D.,
RA Blumberg S., Shoham G.;
RT "Streptomyces griseus aminopeptidase: X-ray crystallographic structure at
RT 1.75-A resolution.";
RL J. Mol. Biol. 265:620-636(1997).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (1.30 ANGSTROMS) IN COMPLEX WITH CALCIUM AND ZINC AND
RP INHIBITORS, COFACTOR, ACTIVITY REGULATION, SUBUNIT, AND DISULFIDE BOND.
RX PubMed=15388919; DOI=10.1107/s0907444904018281;
RA Reiland V., Gilboa R., Spungin-Bialik A., Schomburg D., Shoham Y.,
RA Blumberg S., Shoham G.;
RT "Binding of inhibitory aromatic amino acids to Streptomyces griseus
RT aminopeptidase.";
RL Acta Crystallogr. D 60:1738-1746(2004).
CC -!- FUNCTION: An exopeptidase specific for larger hydrophobic amino acids
CC (especially leucine), no cleavage occurs if the next residue is proline
CC (PubMed:8444149). {ECO:0000269|PubMed:2503378,
CC ECO:0000269|PubMed:8444149, ECO:0000269|PubMed:8665903}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Release of an N-terminal amino acid with a preference for
CC large hydrophobic amino-terminus residues.; EC=3.4.11.24;
CC Evidence={ECO:0000269|PubMed:8444149};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:15388919, ECO:0000269|PubMed:9048953};
CC Note=Binds 1 Ca(2+) per subunit. {ECO:0000269|PubMed:15388919,
CC ECO:0000269|PubMed:9048953};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:15388919, ECO:0000269|PubMed:8444149,
CC ECO:0000269|PubMed:9048953};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:8444149};
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:8444149};
CC Note=Binds 2 Zn(2+) ions per subunit. Can be replaced by Mn(2+) or
CC Co(2+) which results in altered specificities (PubMed:8444149).
CC {ECO:0000269|PubMed:15388919, ECO:0000269|PubMed:8444149,
CC ECO:0000269|PubMed:9048953};
CC -!- ACTIVITY REGULATION: Calcium activates the enzyme, inhibited by 1,10-
CC phenanthroline, EDTA and EGTA (PubMed:2503378). End-product inhibited
CC by L-amino acids (PubMed:15388919). Non-competitively inhibited by NaF
CC and NaH(2)PO(4) (PubMed:17608735). {ECO:0000269|PubMed:15388919,
CC ECO:0000269|PubMed:17608735, ECO:0000269|PubMed:2503378,
CC ECO:0000269|PubMed:8444149}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.59 mM for Leu-NH-Np (with zinc as cofactor)
CC {ECO:0000269|PubMed:8444149};
CC KM=0.13 mM for Leu-NH-Np (with manganese as cofactor)
CC {ECO:0000269|PubMed:8444149};
CC KM=0.046 mM for Leu-NH-Np (with cobalt as cofactor)
CC {ECO:0000269|PubMed:8444149};
CC KM=4.1 mM for Ala-NH-Np (with zinc as cofactor)
CC {ECO:0000269|PubMed:8444149};
CC KM=1.74 mM for Ala-NH-Np (with cobalt as cofactor)
CC {ECO:0000269|PubMed:8444149};
CC KM=0.57 mM for Leu-pNA (with zinc as cofactor)
CC {ECO:0000269|PubMed:15280041};
CC Note=kcat is 390 sec(-1) with Leu-pNA. {ECO:0000269|PubMed:15280041};
CC Temperature dependence:
CC Stable after heating at 69 degrees Celsius for 5 hours.
CC {ECO:0000269|PubMed:2503378};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:2503378,
CC ECO:0000269|PubMed:9048953}.
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:2503378,
CC ECO:0000269|PubMed:8665903}.
CC -!- MASS SPECTROMETRY: Mass=29728; Mass_error=1.0; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:8665903};
CC -!- BIOTECHNOLOGY: Sold by several companies as an extracellular extract
CC called Pronase. {ECO:0000305|PubMed:2503378,
CC ECO:0000305|PubMed:8444149, ECO:0000305|PubMed:8665903}.
CC -!- MISCELLANEOUS: Most experiments use protein purified from a commercial
CC source rather than protein that has been overexpressed. {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the peptidase M28 family. M28A subfamily.
CC {ECO:0000305}.
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DR EMBL; AP009493; BAG22638.1; -; Genomic_DNA.
DR PIR; S66427; S66427.
DR RefSeq; WP_012381549.1; NC_010572.1.
DR PDB; 1CP7; X-ray; 1.58 A; A=46-329.
DR PDB; 1F2O; X-ray; 1.70 A; A=46-329.
DR PDB; 1F2P; X-ray; 1.80 A; A=46-329.
DR PDB; 1QQ9; X-ray; 1.53 A; A=46-329.
DR PDB; 1TF8; X-ray; 1.30 A; A=46-329.
DR PDB; 1TF9; X-ray; 1.30 A; A=46-329.
DR PDB; 1TKF; X-ray; 1.20 A; A=46-329.
DR PDB; 1TKH; X-ray; 1.25 A; A=46-329.
DR PDB; 1TKJ; X-ray; 1.15 A; A=46-329.
DR PDB; 1XBU; X-ray; 1.20 A; A=46-322.
DR PDB; 1XJO; X-ray; 1.75 A; A=46-329.
DR PDBsum; 1CP7; -.
DR PDBsum; 1F2O; -.
DR PDBsum; 1F2P; -.
DR PDBsum; 1QQ9; -.
DR PDBsum; 1TF8; -.
DR PDBsum; 1TF9; -.
DR PDBsum; 1TKF; -.
DR PDBsum; 1TKH; -.
DR PDBsum; 1TKJ; -.
DR PDBsum; 1XBU; -.
DR PDBsum; 1XJO; -.
DR AlphaFoldDB; P80561; -.
DR SMR; P80561; -.
DR STRING; 455632.SGR_5809; -.
DR DrugBank; DB04713; 4-Iodo-D-phenylalanine.
DR DrugBank; DB03660; 4-Iodo-L-phenylalanine.
DR DrugBank; DB02467; L-methionine (S)-S-oxide.
DR MEROPS; M28.003; -.
DR EnsemblBacteria; BAG22638; BAG22638; SGR_5809.
DR GeneID; 6210128; -.
DR KEGG; sgr:SGR_5809; -.
DR PATRIC; fig|455632.4.peg.5952; -.
DR eggNOG; COG2234; Bacteria.
DR eggNOG; COG4935; Bacteria.
DR HOGENOM; CLU_024336_2_0_11; -.
DR OMA; DHASFKN; -.
DR OrthoDB; 1465483at2; -.
DR BRENDA; 3.4.11.24; 6035.
DR SABIO-RK; P80561; -.
DR EvolutionaryTrace; P80561; -.
DR Proteomes; UP000001685; Chromosome.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0004177; F:aminopeptidase activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008235; F:metalloexopeptidase activity; IEA:InterPro.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IEA:InterPro.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR CDD; cd03876; M28_SGAP_like; 1.
DR InterPro; IPR008979; Galactose-bd-like_sf.
DR InterPro; IPR045175; M28_fam.
DR InterPro; IPR041756; M28_SGAP-like.
DR InterPro; IPR002884; P_dom.
DR InterPro; IPR007484; Peptidase_M28.
DR PANTHER; PTHR12147; PTHR12147; 1.
DR Pfam; PF01483; P_proprotein; 1.
DR Pfam; PF04389; Peptidase_M28; 1.
DR SUPFAM; SSF49785; SSF49785; 1.
DR PROSITE; PS51829; P_HOMO_B; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Aminopeptidase; Calcium; Direct protein sequencing;
KW Disulfide bond; Hydrolase; Metal-binding; Protease; Secreted; Signal; Zinc;
KW Zymogen.
FT SIGNAL 1..45
FT /evidence="ECO:0000269|PubMed:2503378,
FT ECO:0000269|PubMed:8665903"
FT CHAIN 46..329
FT /note="Aminopeptidase S"
FT /id="PRO_0000174143"
FT PROPEP 330..445
FT /note="Removed in mature form"
FT /evidence="ECO:0000269|PubMed:8665903"
FT /id="PRO_0000431389"
FT DOMAIN 325..445
FT /note="P/Homo B"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01173"
FT ACT_SITE 176
FT /note="Proton acceptor"
FT /evidence="ECO:0000303|PubMed:15280041,
FT ECO:0000303|PubMed:17608735"
FT BINDING 48
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT BINDING 49
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT BINDING 130
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT BINDING 142
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT BINDING 142
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT BINDING 177
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT BINDING 205
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT BINDING 292
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /ligand_note="catalytic"
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT BINDING 307
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT BINDING 311
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT SITE 291
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000303|PubMed:15280041,
FT ECO:0000303|PubMed:17608735"
FT DISULFID 290..295
FT /evidence="ECO:0000269|PubMed:15388919,
FT ECO:0000269|PubMed:9048953"
FT MUTAGEN 176
FT /note="E->A: KM halves (0.33 mM), kcat reduces by 5 orders
FT of magnitude."
FT /evidence="ECO:0000269|PubMed:15280041"
FT MUTAGEN 176
FT /note="E->D,Q: KM decreases (to 0.4 mM), kcat reduces by 4
FT orders of magnitude."
FT /evidence="ECO:0000269|PubMed:15280041"
FT MUTAGEN 291
FT /note="Y->A,F,S: Activity decreases about 100-fold."
FT /evidence="ECO:0000269|PubMed:15280041"
FT CONFLICT 185
FT /note="Y -> F (in Ref. 2; AA sequence)"
FT CONFLICT 285
FT /note="Q -> R (in Ref. 5; no nucleotide entry)"
FT HELIX 51..66
FT /evidence="ECO:0007829|PDB:1TKJ"
FT TURN 67..70
FT /evidence="ECO:0007829|PDB:1TKJ"
FT HELIX 77..93
FT /evidence="ECO:0007829|PDB:1TKJ"
FT STRAND 96..104
FT /evidence="ECO:0007829|PDB:1TKJ"
FT STRAND 107..115
FT /evidence="ECO:0007829|PDB:1TKJ"
FT STRAND 119..130
FT /evidence="ECO:0007829|PDB:1TKJ"
FT TURN 140..143
FT /evidence="ECO:0007829|PDB:1TKJ"
FT HELIX 144..159
FT /evidence="ECO:0007829|PDB:1TKJ"
FT STRAND 165..174
FT /evidence="ECO:0007829|PDB:1TKJ"
FT HELIX 176..178
FT /evidence="ECO:0007829|PDB:1TKJ"
FT HELIX 181..189
FT /evidence="ECO:0007829|PDB:1TKJ"
FT HELIX 192..195
FT /evidence="ECO:0007829|PDB:1TKJ"
FT STRAND 198..204
FT /evidence="ECO:0007829|PDB:1TKJ"
FT STRAND 215..217
FT /evidence="ECO:0007829|PDB:1TKF"
FT HELIX 221..233
FT /evidence="ECO:0007829|PDB:1TKJ"
FT STRAND 239..241
FT /evidence="ECO:0007829|PDB:1TKF"
FT HELIX 250..255
FT /evidence="ECO:0007829|PDB:1TKJ"
FT STRAND 260..264
FT /evidence="ECO:0007829|PDB:1TKJ"
FT STRAND 268..270
FT /evidence="ECO:0007829|PDB:1TKJ"
FT HELIX 273..279
FT /evidence="ECO:0007829|PDB:1TKJ"
FT STRAND 285..287
FT /evidence="ECO:0007829|PDB:1TKJ"
FT TURN 289..292
FT /evidence="ECO:0007829|PDB:1TKJ"
FT HELIX 303..321
FT /evidence="ECO:0007829|PDB:1TKJ"
SQ SEQUENCE 445 AA; 45940 MW; 6B42B63191F700C9 CRC64;
MRPNRFSLRR SPTAVAAVAL AAVLAAGAPA AQAAGAAAPT AAAAAAPDIP LANVKAHLTQ
LSTIAANNGG NRAHGRPGYK ASVDYVKAKL DAAGYTTTLQ QFTSGGATGY NLIADWPGGD
PNKVLMAGAH LDSVSSGAGI NDNGSGSAAV LETALAVSRA GYQPDKHLRF AWWGAEELGL
IGSKYYVNNL PSADRSKLAG YLNFDMIGSP NPGYFVYDDD PVIEKTFKDY FAGLNVPTEI
ETEGDGRSDH APFKNVGVPV GGLFTGAGYT KSAAQAQKWG GTAGQAFDRC YHSSCDSLSN
INDTALDRNS DAAAHAIWTL SSGTGEPPTG EGVFSNTTDV AIPDAGAAVT SSVAVTGRTG
NAPAALQVGV DIKHTYRGDL VVDLLAPDGT AYRLKNSSSG DSADNVIATY TVNASSEVAN
GSWKLRVQDI ARQDTGYIDS WKLTF
