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Sequencing, expression and biochemical characterization of the Porphyromonas gingivalis pepO gene encoding a protein homologous to human endothelin‐converting enzyme
Author(s) -
Awano Shuji,
Ansai Toshihiro,
Mochizuki Hajime,
Yu Weixian,
Tanzawa Kazuhiko,
Turner Anthony J.,
Takehara Tadamichi
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)01326-5
Subject(s) - phosphoramidon , porphyromonas gingivalis , microbiology and biotechnology , recombinant dna , nucleic acid sequence , peptide sequence , biology , open reading frame , complementary dna , gene , sequence analysis , homology (biology) , biochemistry , enzyme , amino acid , genetics , bacteria , neprilysin
We have determined the nucleotide sequence of the clone pAL2 obtained from Porphyromonas gingivalis 381 in the previous study [Ansai et al. (1995) Microbiology 141, 2047–2052]. The DNA sequence analysis of this fragment revealed one complete ORF and one incomplete ORF. The ORF encoded a protein (PgPepO) of 690 amino acids with a calculated molecular weight of 78 796. The deduced amino acid sequence exhibited a significant homology with human endothelin‐converting enzyme (ECE)‐1. Recombinant PgPepO was purified to homogeneity and characterized. The purified enzyme was strongly inhibited by phosphoramidon, and converted big endothelin‐1 to endothelin‐1. Furthermore, the purified PgPepO strongly cross‐reacted with a monoclonal antibody against rat ECE‐1. These results indicate that PgPepO has striking similarity to mammalian ECE in structure and function.