cDNA cloning and primary structure of tryptase from bovine mast cells, and evidence for the expression of bovine pancreatic trypsin inhibitor mRNA in the same cells.
A partial cDNA encoding bovine tryptase, an oligomeric serine proteinase previously isolated from bovine mast cells, was obtained by reverse transcription/polymerase chain reaction of mast cell mRNA, using combinations of primers designed on the basis of information obtained from partial sequencing of the purified protein. The complete amino acid sequence of bovine tryptase (245 residues) was deduced from a 711-bp nucleotide sequence and from Edman degradation of the protein. Bovine tryptase primary structure has an identity of about 75% with tryptases from other species and includes all the essential residues of the active-site regions; sequence data in the region of the putative substrate binding pocket suggest a rearrangement capable of maintaining the specificity of trypsin-like proteinases. From the same mast cell mRNA, cDNA encoding bovine trypsin protease inhibitor (BPTI) was obtained and amplified with specific primers, confirming the synthesis of BPTI in these cells. Results are consistent with previous data on the presence of BPTI and bovine tryptase in the same granules of bovine mast cells and with their interaction in vitro.
Pallaoro
M
M
Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università Tor Vergata, Roma, Italia.
Gambacurta
A
A
Fiorucci
L
L
Mignogna
G
G
Barra
D
D
Ascoli
F
F
D000595:Amino Acid Sequence; D000818:Animals; D007611:Aprotinin / Q000235:genetics*; D001483:Base Sequence; D002417:Cattle; D053818:Chymases; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006801:Humans; D008407:Mast Cells / Q000201:enzymology*; D008969:Molecular Sequence Data; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics*; D053802:Tryptases
D018076:DNA, Complementary; D007611:Aprotinin; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases