LOCUS CDQ48116.1 918 aa PRT BCT 04-FEB-2016 DEFINITION Vibrio anguillarum EmpA processing protease; protein. ACCESSION LK021129-219 PROTEIN_ID CDQ48116.1 SOURCE Vibrio anguillarum ORGANISM Vibrio anguillarum Bacteria; Proteobacteria; Gammaproteobacteria; Vibrionales; Vibrionaceae; Vibrio. REFERENCE 1 (bases 1 to 1187342) AUTHORS Holm K. JOURNAL Submitted (26-MAR-2014) to the INSDC. Norstruct, Dept of Chemistry, University of Tromso, Science Park 3, NO-9037 Tromso, NORWAY. REFERENCE 2 AUTHORS Holm K.O., Nilsson K., Hjerde E., Willassen N.P., Milton D.L. TITLE Complete genome sequence of Vibrio anguillarum strain NB10, a virulent isolate from the Gulf of Bothnia JOURNAL Stand Genomic Sci 10, 60-60(2015). PUBMED 26380645 FEATURES Qualifiers source /organism="Vibrio anguillarum" /chromosome="2" /host="Rainbow trout" /strain="NB10" /mol_type="genomic DNA" /country="Sweden:Baltic Sea, Norrbyn Umeaa" /isolation_source="clinical isolate, Rainbow trout" /serovar="O1" /db_xref="taxon:55601" protein /transl_table=11 /gene="empA-p" /gene_synonym="epp" /locus_tag="VANGNB10_cII0219c" /old_locus_tag="TVA2585" /function="Ref: The zinc metalloprotease EmpA is a virulence factor for the fish pathogen Vibrio anguillarum. Previous studies demonstrated that EmpA is secreted as a 46-kDa proenzyme that is activated extracellularly by the removal of an 10-kDa propeptide. We hypothesized that a specific protease is responsible for processing secreted pro-EmpA into mature EmpA. To identify the protease responsible for processing pro-EmpA, a minitransposon mutagenesis (using mini-Tn10Km) clone bank of V. anguillarum was screened for reduced protease activity due to insertions in undescribed genes. One mutant with reduced protease activity was identified. The region containing the mini-Tn10Km was cloned, sequenced, and found to contain epp, an open reading frame encoding a putative protease. Further characterization of epp was done using strain M101,created by single-crossover insertional mutagenesis. Protease activity was absent in M101 cultures even when empA protease activity was induced by salmon gastrointestinal mucus. When the epp mutation was complemented with a wild-type copy of epp (M102), protease activity was restored. Western blot analysis of sterile filtered culture supernatants from wild-type (M93Sm) cells,M101 cells, and M102 cells revealed that only pro-EmpA was present in M101supernatants; both pro-EmpA and mature EmpA were detected in M93Sm and M102 supernatants. When sterile filtered culture supernatants from the empA mutant strain (M99) and M101 were mixed, protease activity was restored. Western blot analysis revealed that pro-EmpA in M101 culture supernatant was processed to mature EmpA only after mixing with M99 culture supernatant. These data show that Epp is the EmpA-processing protease." /note="user locus_tag: VANGcII0219c" /note="Similar to Vibrio anguillarum (Listonella anguillarum) epp subname: full=empa processing protease UniProt:B5LBF9 (EMBL:EU650390) (918 aa) fasta scores: E()=0, 99.3% id in 918 aa, and to Vibrio anguillarum (Listonella anguillarum) epp subname: full=empa processing protease UniProt:B5LBF9 (EMBL:EU650390) (918 aa) fasta scores: E()=0, 99.3% id in 918 aa" BEGIN 1 MKHIRKTLLA SAMVTLFNVS AYAHTPIDLG VVNEDKLIEM LVRNGVVDAN ATDAIKRDAL 61 DRYLEQKIKS GFHGDAQFGK QALEQRARVL KTIEKQKGHQ KASVFALDIS TKRTDKVLAL 121 LIDFPDLPWN DNRLTKEHTE MLYASYNPPH YQDLLFSDAG YAGPNGENFI SMRQYYESES 181 GQSYSVAGQA AGWYRASKNA AFYGGNSATT DKDMNAQELV REALNQLAQD PSINLADYDI 241 EDRYDYNGNG NFREPDGVID HLMVFHASVG EEAGGGVLGP DAIWSHRYNL GQPYVLQGTK 301 STLPDRFNGQ YAAFDYTMQP IDAAAGVCAH EYGHDLGLPD EYDTQYTGKG EPVSYWSIMS 361 SGSWAGKIGG TQPTAFSSWS KQFLQKAVGG RWINDDQISI AELEKNPRVF TLFQTTDNAR 421 PNMVKIDLPL KRVDGVKPYQ GSSAFYSNRG DDLKNSMSRK LSIPTGEQAT LNFKAWYQIE 481 KDYDFARVLI NGKAIAGNIT TMDDPYNTGL VPAIAGQSDG WVDAIFDVSQ WAGQEVTLSF 541 DYVTDGGLAM EGLYLDNLQL EVDGQQTLID DAEGTSTFAF KGYTTNNGFH EAPHYYLLQW 601 RSHNDVDQGL ANIKRMGQLM TFEPGLLVWY VDESYTDNWV GNHPGEGWLG VVDADQNALV 661 WSKTGEAAQT RFQVRDATFS LQDQTPLRLV AADGDVLEDT SLIGNGNFSD GADYSSPLVP 721 DAGRKLTEFG LMIDILNQGQ DNEYGVIRLS KVSEQNQAPV ANFELKTDGL SVSAFNYSYD 781 EDGELVSYAW DFGNGQMSSE MAPSWSYTKA GQYTVSLTVT DDKGATNTTT RTTQVEVPNA 841 LPQASAKYIH LGRWVTMWST SSDSDGRIVD TEWTLPNGKI KRGKLFTAIF PSSGKHEVKI 901 KVMDDKGATV TNTIVVNL //