Purification and molecular cloning of chymase from human tonsils

FEBS Letters

Enomoto, Mitsuo¹  Katunuma, Nobuhiko²  Sukenaga, Yoshikazu¹  Kido, Hiroshi²  Neki, Akio¹  Ishida, Koichi¹  Takagi, Kenkichi¹ 

[1] Research Laboratories Pharmaceuticals Group, Nippon Kayaku Co., Kita-ku, Tokyo 114, Japan;Department of Enzyme Chemistry, Institute for Enzyme Research, School of Medicine, The University of Tokushima, Tokushima 770, Japan

关键词: Chymase;    Human tonsil;    cDNA cloning;    Amino acid sequence;    HPLC;    high performance liquid chromatography;    Suc;    Succinyl;    MCA;    4-methylcoumaryl-7-amide;    PCR;    polymerase chain reaction;    bp;    base pair;    kDa;    kilodaltons;    SDS-PAGE;    sodium dodecyl polyacrylamide gel electrophoresis;    TFA;    trifluoroacetic acid;   

DOI  :  10.1016/0014-5793(93)81461-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown