caa a22 4500 397556977 CHVBK 20180308164805.0 cr unu---uuuuu 161202e199608 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021426 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021426 Cloning and Sequencing of the Gene Encoding a Novel Lysine-Specific Cysteine Proteinase (Lys-Gingipain) in Porphyromonas gingivalis: Structural Relationship with the Arginine-Specific Cysteine Proteinase (Arg-Gingipain) [Elektronische Daten] [Kuniaki Okamoto, Tomoko Kadowaki, Koji Nakayama, Kenji Yamamoto] Lys-gingipain (KGP), so termed due to its peptide cleavage specificity for lysine residues, is a cysteine proteinase produced by the Gram-negative anaerobic bacterium Porphyromonas gingivalis. Mixed oligonucleotide primers designed from the NH2-terminal sequence of the purified enzyme were used to clone the KGP-encoding gene (kgp) from the organism. The nucleotide sequence of kgp had a 5,169-bp open reading frame encoding 1,723 amino acids with a calculated molecular mass of 218 kDa. As the extracellular mature enzyme had an apparent molecular mass of 51 kDa in gels, the precursor of KGP was found to comprise at least four domains, the signal peptide, the NH2-terminal prodomain, the mature proteinase domain, and the COOH-terminal hemagglutinin domain, and to be proteolytically processed during its transport. Importantly, the COOH-terminal region contained three direct repeats of two different amino acid sequences, LKWD(or E)AP and YTYTVYRDGTKI, and the subdomains located between the two repeats exhibited strong similarity to those of Arg-gingipain (RGP), another major cysteine proteinase produced by the organism and having cleavage specificity for arginine residues, although the arrangement of the subdomains was not necessarily identical in the two enzymes. Since the KGP activity was greatly decreased in RGP-deficient mutants and since the most probable site of the propeptide cleavage was present in the homologous sequence highly susceptible to proteolysis by RGP, the precursor of KGP is likely to be processed by RGP to form the mature enzyme. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence Arg-gingipain nationallicence Lys-gingipain nationallicence lysine-specific cysteine proteinase nationallicence Porphyromonas gingivalis nationallicence precursor structure nationallicence Okamoto Kuniaki Department of Pharmacology Kyushu University Faculty of Dentistry Higashi-ku, Fukuoka 812-82 aut Kadowaki Tomoko Department of Pharmacology Kyushu University Faculty of Dentistry Higashi-ku, Fukuoka 812-82 aut Nakayama Koji Department of Microbiology Kyushu University Faculty of Dentistry Higashi-ku, Fukuoka 812-82 aut Yamamoto Kenji Department of Pharmacology Kyushu University Faculty of Dentistry Higashi-ku, Fukuoka 812-82 aut The Journal of Biochemistry Oxford University Press 120/2(1996-08), 398-406 0021-924X 120:2<398 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021426 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021426 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Okamoto Kuniaki Department of Pharmacology Kyushu University Faculty of Dentistry Higashi-ku, Fukuoka 812-82 aut NATIONALLICENCE 700 1- Kadowaki Tomoko Department of Pharmacology Kyushu University Faculty of Dentistry Higashi-ku, Fukuoka 812-82 aut NATIONALLICENCE 700 1- Nakayama Koji Department of Microbiology Kyushu University Faculty of Dentistry Higashi-ku, Fukuoka 812-82 aut NATIONALLICENCE 700 1- Yamamoto Kenji Department of Pharmacology Kyushu University Faculty of Dentistry Higashi-ku, Fukuoka 812-82 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/2(1996-08), 398-406 0021-924X 120:2<398 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford