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Purification and molecular cloning of chymase from human tonsils
Author(s) -
Sukenaga Yoshikazu,
Kido Hiroshi,
Neki Akio,
Enomoto Mitsuo,
Ishida Koichi,
Takagi Kenkichi,
Katunuma Nobuhiko
Publication year - 1993
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 - 10.1016/0014-5793(93)81461-8
Subject(s) - chymase , microbiology and biotechnology , complementary dna , molecular mass , molecular cloning , peptide sequence , biology , biochemistry , protease , enzyme , cloning (programming) , amino acid , primer (cosmetics) , protein primary structure , chemistry , gene , organic chemistry , computer science , programming language
A chymotrypsin‐like protease was purified to homogeneity from human tonsils by a series of Chromatographie procedures. The purified enzyme gave a single protein band with an apparent molecular mass of 30 kDa on SDS‐PAGE. The sequence of the first 21 amino acids at the N‐terminus of the enzyme was determined. A cDNA for the enzyme was cloned by PCR amplification from extracted tonsillar mRNA using a supposed N‐terminal oligonucleotide primer and a conserved C‐terminal primer of the chymase family. The deduced amino acid sequence of the isolated clone was identical to that of human chymase in connective tissue‐type mast cells from heart except for a Ser instead of a Cys at the N‐terminal 7th position.