Purification and molecular cloning of chymase from human tonsils.
A chymotrypsin-like protease was purified to homogeneity from human tonsils by a series of chromatographic 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.
Research Laboratories Pharmaceuticals Group, Nippon Kayaku Co., Tokyo, Japan.
Y
Y
Sukenaga
H
H
Kido
A
A
Neki
M
M
Enomoto
K
K
Ishida
K
K
Takagi
N
N
Katunuma
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002648:Child; D053818:Chymases; D003001:Cloning, Molecular; D004247:DNA; D006801:Humans; D008969:Molecular Sequence Data; D014066:Palatine Tonsil / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000235:genetics* / Q000302:isolation & purification
D004247:DNA; D012697:Serine Endopeptidases; D053818:Chymases