ID ASP_AERSO Reviewed; 624 AA. AC Q9L5A4; DT 14-DEC-2022, integrated into UniProtKB/Swiss-Prot. DT 01-SEP-2009, sequence version 2. DT 14-DEC-2022, entry version 107. DE RecName: Full=Aeromonas extracellular serine protease {ECO:0000303|PubMed:12153115}; DE Short=ASP {ECO:0000303|PubMed:12446656}; DE EC=3.4.21.121 {ECO:0000269|PubMed:16487335, ECO:0000269|PubMed:18067862, ECO:0000269|PubMed:18462393, ECO:0000269|PubMed:19654332}; DE AltName: Full=Lys-Lys/Arg-Xaa endopeptidase {ECO:0000305}; DE Flags: Precursor; GN Name=asp {ECO:0000303|PubMed:18462393}; OS Aeromonas sobria. OC Bacteria; Proteobacteria; Gammaproteobacteria; Aeromonadales; OC Aeromonadaceae; Aeromonas. OX NCBI_TaxID=646; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 25-34, FUNCTION, RP ACTIVITY REGULATION, AND SUBCELLULAR LOCATION. RC STRAIN=288; RX PubMed=11092244; DOI=10.1111/j.1348-0421.2000.tb02565.x; RA Okamoto K., Nomura T., Hamada M., Fukuda T., Noguchi Y., Fujii Y.; RT "Production of serine protease of Aeromonas sobria is controlled by the RT protein encoded by the gene lying adjacent to the 3' end of the protease RT gene."; RL Microbiol. Immunol. 44:787-798(2000). RN [2] RP FUNCTION, ACTIVITY REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES. RC STRAIN=288; RX PubMed=12153115; DOI=10.1111/j.1348-0421.2002.tb02710.x; RA Yokoyama R., Fujii Y., Noguchi Y., Nomura T., Akita M., Setsu K., RA Yamamoto S., Okamoto K.; RT "Physicochemical and biological properties of an extracellular serine RT protease of Aeromonas sobria."; RL Microbiol. Immunol. 46:383-390(2002). RN [3] RP ACTIVITY REGULATION, INTERACTION WITH ORF2, AND SUBCELLULAR LOCATION. RX PubMed=12446656; DOI=10.1128/jb.184.24.7058-7061.2002; RA Nomura T., Fujii Y., Yamanaka H., Kobayashi H., Okamoto K.; RT "The protein encoded at the 3' end of the serine protease gene of Aeromonas RT sobria functions as a chaperone in the production of the protease."; RL J. Bacteriol. 184:7058-7061(2002). RN [4] RP FUNCTION, AND CATALYTIC ACTIVITY. RC STRAIN=288; RX PubMed=16487335; DOI=10.1111/j.1574-6968.2006.00134.x; RA Kobayashi H., Takahashi E., Oguma K., Fujii Y., Yamanaka H., Negishi T., RA Arimoto-Kobayashi S., Tsuji T., Okamoto K.; RT "Cleavage specificity of the serine protease of Aeromonas sobria, a member RT of the kexin family of subtilases."; RL FEMS Microbiol. Lett. 256:165-170(2006). RN [5] RP FUNCTION. RC STRAIN=T94; RX PubMed=17142774; DOI=10.4049/jimmunol.177.12.8723; RA Imamura T., Kobayashi H., Khan R., Nitta H., Okamoto K.; RT "Induction of vascular leakage and blood pressure lowering through kinin RT release by a serine proteinase from Aeromonas sobria."; RL J. Immunol. 177:8723-8729(2006). RN [6] RP FUNCTION, AND CATALYTIC ACTIVITY. RX PubMed=18067862; DOI=10.1016/j.febslet.2007.11.076; RA Nitta H., Kobayashi H., Irie A., Baba H., Okamoto K., Imamura T.; RT "Activation of prothrombin by ASP, a serine protease released from RT Aeromonas sobria."; RL FEBS Lett. 581:5935-5939(2007). RN [7] RP FUNCTION, CATALYTIC ACTIVITY, SUBCELLULAR LOCATION, AND DISRUPTION RP PHENOTYPE. RC STRAIN=288; RX PubMed=18462393; DOI=10.1111/j.1574-6968.2008.01184.x; RA Imamura T., Nitta H., Wada Y., Kobayashi H., Okamoto K.; RT "Impaired plasma clottability induction through fibrinogen degradation by RT ASP, a serine protease released from Aeromonas sobria."; RL FEMS Microbiol. Lett. 284:35-42(2008). RN [8] RP FUNCTION. RX PubMed=18714034; DOI=10.4049/jimmunol.181.5.3602; RA Nitta H., Imamura T., Wada Y., Irie A., Kobayashi H., Okamoto K., Baba H.; RT "Production of C5a by ASP, a serine protease released from Aeromonas RT sobria."; RL J. Immunol. 181:3602-3608(2008). RN [9] RP ACTIVITY REGULATION. RX PubMed=23089609; DOI=10.1515/hsz-2012-0117; RA Murakami Y., Wada Y., Kobayashi H., Irie A., Hasegawa M., Yamanaka H., RA Okamoto K., Eto M., Imamura T.; RT "Inhibition of Aeromonas sobria serine protease (ASP) by alpha2- RT macroglobulin."; RL Biol. Chem. 393:1193-1200(2012). RN [10] {ECO:0007744|PDB:3HJR} RP X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) OF 25-624 IN COMPLEX WITH CALCIUM, RP SEQUENCE REVISION, IDENTIFICATION BY MASS SPECTROMETRY, FUNCTION, CATALYTIC RP ACTIVITY, COFACTOR, SUBCELLULAR LOCATION, DOMAIN, AND DISULFIDE BOND. RC STRAIN=T94; RX PubMed=19654332; DOI=10.1074/jbc.m109.006114; RA Kobayashi H., Utsunomiya H., Yamanaka H., Sei Y., Katunuma N., Okamoto K., RA Tsuge H.; RT "Structural basis for the kexin-like serine protease from Aeromonas sobria RT as sepsis-causing factor."; RL J. Biol. Chem. 284:27655-27663(2009). RN [11] {ECO:0007744|PDB:3WQB} RP X-RAY CRYSTALLOGRAPHY (1.41 ANGSTROMS) OF 25-624 IN COMPLEX WITH ORF2 AND RP CALCIUM, DISULFIDE BONDS, FUNCTION, COFACTOR, ACTIVITY REGULATION, RP INTERACTION WITH ORF2, AND SUBCELLULAR LOCATION. RX PubMed=25784551; DOI=10.1074/jbc.m114.622852; RA Kobayashi H., Yoshida T., Miyakawa T., Tashiro M., Okamoto K., Yamanaka H., RA Tanokura M., Tsuge H.; RT "Structural basis for action of the external chaperone for a propeptide- RT deficient serine protease from Aeromonas sobria."; RL J. Biol. Chem. 290:11130-11143(2015). CC -!- FUNCTION: Exhibits serine protease activity (PubMed:11092244, CC PubMed:12153115, PubMed:16487335, PubMed:18067862, PubMed:18462393, CC PubMed:19654332). Preferentially cleaves the peptide bond following two CC basic residues, one of which is Lys, but does not recognize the bond CC following a single basic residue (PubMed:16487335). Probable potent CC virulence factor that cleaves various host plasma proteins, including CC prekallikrein, prothrombin and fibrinogen (PubMed:16487335, CC PubMed:17142774, PubMed:18067862, PubMed:18462393, PubMed:19654332). CC ASP induces vascular leakage and reduction in blood pressure by CC activating the host plasma kallikrein/kinin system (PubMed:12153115, CC PubMed:16487335, PubMed:17142774). It affects the host coagulation CC system during infection through activation of prothrombin to alpha- CC thrombin and degradation of fibrinogen, which impairs plasma CC clottability (PubMed:18067862, PubMed:18462393). It also hydrolyzes the CC complement component C5, releasing the C5a anaphylatoxin, which causes CC the formation of pus and edema (PubMed:18714034). In addition, degrades CC its external chaperone ORF2 after the secretion of the ASP-ORF2 complex CC (PubMed:25784551). {ECO:0000269|PubMed:11092244, CC ECO:0000269|PubMed:12153115, ECO:0000269|PubMed:16487335, CC ECO:0000269|PubMed:17142774, ECO:0000269|PubMed:18067862, CC ECO:0000269|PubMed:18462393, ECO:0000269|PubMed:18714034, CC ECO:0000269|PubMed:19654332, ECO:0000269|PubMed:25784551}. CC -!- CATALYTIC ACTIVITY: CC Reaction=Cleavage of -Lys-Lys-|-Xaa and -Lys-Arg-|-Xaa bonds.; CC EC=3.4.21.121; Evidence={ECO:0000269|PubMed:16487335, CC ECO:0000269|PubMed:18067862, ECO:0000269|PubMed:18462393, CC ECO:0000269|PubMed:19654332}; CC -!- COFACTOR: CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108; CC Evidence={ECO:0000305|PubMed:19654332, ECO:0000305|PubMed:25784551}; CC Note=Binds 3 calcium ions per subunit. {ECO:0000269|PubMed:19654332, CC ECO:0000269|PubMed:25784551}; CC -!- ACTIVITY REGULATION: Folding, maturation and production of the active CC form of the protease by the cell requires a protein (ORF2), encoded CC just downstream of asp, which acts as a chaperone (PubMed:11092244, CC PubMed:12446656, PubMed:25784551). Formation of a complex with ORF2 in CC the periplasm also inactivates the protease activity and likely CC protects ASP from intrinsic proteases (PubMed:25784551). In vitro, CC protease activity is inhibited by human alpha-2-macroglobulin, CC suggesting that this inhibitor can impede ASP virulence activities in CC A.sobria infection sites (PubMed:23089609). However, slow ASP CC inhibition by alpha-2-macroglobulin in plasma may indicate insufficient CC ASP control in vivo (PubMed:23089609). Activity is inhibited by serine CC protease inhibitors such as 4-(2-aminoethyl)-benzenesulfonyl fluoride CC (AEBSF) and diisopropyl fluorophosphate (DFP) (PubMed:11092244, CC PubMed:12153115). Not inhibited by metallo-protease inhibitors and CC cysteine protease inhibitors (PubMed:12153115). The treatment with CC reagents to modify sulfhydryl group do not reduce the activity CC (PubMed:12153115). {ECO:0000269|PubMed:11092244, CC ECO:0000269|PubMed:12153115, ECO:0000269|PubMed:12446656, CC ECO:0000269|PubMed:23089609, ECO:0000269|PubMed:25784551}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC pH dependence: CC Optimum pH is 7.5. {ECO:0000269|PubMed:12153115}; CC -!- SUBUNIT: Forms a complex with the chaperone ORF2 in the periplasm CC (PubMed:12446656, PubMed:25784551). After translocation of the ASP-ORF2 CC complex from the periplasm to the extracellular space, the complex is CC dissociated in a pH-dependent manner (PubMed:25784551). CC {ECO:0000269|PubMed:12446656, ECO:0000269|PubMed:25784551}. CC -!- SUBCELLULAR LOCATION: Periplasm {ECO:0000269|PubMed:12446656, CC ECO:0000269|PubMed:25784551}. Secreted {ECO:0000269|PubMed:11092244, CC ECO:0000269|PubMed:18462393, ECO:0000269|PubMed:19654332, CC ECO:0000269|PubMed:25784551}. Note=Translocated from the periplasm to CC the extracellular space as a complex with ORF2 (PubMed:25784551). CC Secreted at the infection sites or in the circulation CC (PubMed:18462393). {ECO:0000269|PubMed:18462393, CC ECO:0000269|PubMed:25784551}. CC -!- DOMAIN: Contains an N-terminal subtilisin domain and a C-terminal P- CC domain. {ECO:0000269|PubMed:19654332}. CC -!- DISRUPTION PHENOTYPE: Disruption mutant releases negligible serine CC protease activity and does not affect plasma clotting. CC {ECO:0000269|PubMed:18462393}. CC -!- SIMILARITY: Belongs to the peptidase S8 family. Furin subfamily. CC {ECO:0000305}. 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; AF253471; AAF64521.2; -; Genomic_DNA. DR PDB; 3HJR; X-ray; 1.65 A; A=25-624. DR PDB; 3WQB; X-ray; 1.41 A; A=25-624. DR PDBsum; 3HJR; -. DR PDBsum; 3WQB; -. DR AlphaFoldDB; Q9L5A4; -. DR SMR; Q9L5A4; -. DR MEROPS; S08.125; -. DR KEGG; ag:AAF64521; -. DR BioCyc; MetaCyc:MON-17932; -. DR BRENDA; 3.4.21.121; 169. DR EvolutionaryTrace; Q9L5A4; -. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0004252; F:serine-type endopeptidase activity; IEA:InterPro. DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW. DR CDD; cd04059; Peptidases_S8_Protein_convertases_Kexins_Furin-like; 1. DR Gene3D; 3.40.50.200; -; 1. DR InterPro; IPR008979; Galactose-bd-like_sf. DR InterPro; IPR034182; Kexin/furin. DR InterPro; IPR002884; P_dom. DR InterPro; IPR000209; Peptidase_S8/S53_dom. DR InterPro; IPR036852; Peptidase_S8/S53_dom_sf. DR InterPro; IPR023827; Peptidase_S8_Asp-AS. DR InterPro; IPR022398; Peptidase_S8_His-AS. DR InterPro; IPR023828; Peptidase_S8_Ser-AS. DR InterPro; IPR015500; Peptidase_S8_subtilisin-rel. DR Pfam; PF01483; P_proprotein; 1. DR Pfam; PF00082; Peptidase_S8; 1. DR PRINTS; PR00723; SUBTILISIN. DR SUPFAM; SSF49785; Galactose-binding domain-like; 1. DR SUPFAM; SSF52743; Subtilisin-like; 1. DR PROSITE; PS51829; P_HOMO_B; 1. DR PROSITE; PS51892; SUBTILASE; 1. DR PROSITE; PS00136; SUBTILASE_ASP; 1. DR PROSITE; PS00137; SUBTILASE_HIS; 1. DR PROSITE; PS00138; SUBTILASE_SER; 1. PE 1: Evidence at protein level; KW 3D-structure; Calcium; Direct protein sequencing; Disulfide bond; KW Hydrolase; Metal-binding; Periplasm; Protease; Secreted; Serine protease; KW Signal; Virulence. FT SIGNAL 1..24 FT /evidence="ECO:0000269|PubMed:11092244" FT CHAIN 25..624 FT /note="Aeromonas extracellular serine protease" FT /id="PRO_5004329932" FT DOMAIN 59..421 FT /note="Peptidase S8" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240" FT DOMAIN 456..622 FT /note="P/Homo B" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01173" FT ACT_SITE 102 FT /note="Charge relay system" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240" FT ACT_SITE 139 FT /note="Charge relay system" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240" FT ACT_SITE 360 FT /note="Charge relay system" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240" FT BINDING 53 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="1" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 111 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="1" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 150 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="1" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 152 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="1" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 154 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="1" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 156 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="1" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 321 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="2" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 322 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="2" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 324 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="2" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 327 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="2" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 330 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="2" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 350 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="2" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 478 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="3" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 512 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="3" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 577 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="3" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 579 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="3" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 602 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="3" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT BINDING 603 FT /ligand="Ca(2+)" FT /ligand_id="ChEBI:CHEBI:29108" FT /ligand_label="3" FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT DISULFID 28..48 FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" FT DISULFID 325..350 FT /evidence="ECO:0000269|PubMed:19654332, FT ECO:0000269|PubMed:25784551, ECO:0007744|PDB:3HJR, FT ECO:0007744|PDB:3WQB" SQ SEQUENCE 624 AA; 66635 MW; 6B6F2DC8DE639531 CRC64; MKQTSLALAI TALLSTLPSA LVQANEGCAP LTGKESGMDI GRSSTERCLP GANPLQDQQW YLLNSGQDGF SARGGIAGND LNLWWAHRTG VLGQGVNVAV VDDGLAIAHP DLADNVRPGS KNVVTGSDDP TPTDPDTAHG TSVSGIIAAV DNAIGTKGIA PRAQLQGFNL LDDNSQQLQK DWLYALGDSN ASRDNRVFNQ SYGMSVVDPR SANSLDQSQL DRLFEQQTLK AQGAAYIKAA GNGFNKIAAG GYVLNRTGNG PKLPFENSNL DPSNSNFWNL VVSALNADGV RSSYSSVGSN IFLSATGGEY GTDTPAMVTT DLPGCDMGYN RTDDPSTNRL HGNSQLDASC DYNGVMNGTS SATPSTSGAM ALLMSAYPDL SVRDLRDLLA RSATRVDAKH QPVMVSYTSS TGKVRDVKGL EGWERNAAGM WFSPTYGFGL IDVNKALELA ANHQPLPPLV QLPWQKINVT GSAAAIADVG NSPTSSTTRI ATPLTVEAVQ VMVSLDHQRL PDLLIELVSP AGTRSILLSP FNSLVGQSLD QQQLGFVRTK GLRDMRMLSN KFYGESAQGT WRLEVTDVAN GTRQVSLLNR ETRERTTLTE RNNRQPGKLI SWSLRVLGHD ANRS //