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., Gambacurta, A., Fiorucci, L., Mignogna, G., Barra, D., Ascoli, F. (1996). 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. EUROPEAN JOURNAL OF BIOCHEMISTRY, 237(1), 100-105 [10.1111/j.1432-1033.1996.0100t.x].

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

Gambacurta, A;Fiorucci, L;
1996

Abstract

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.
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore BIO/10
English
tryptase
nucleotide sequence
protein primary structure
bovine mast cells
bovine pancreatic trypsin inhibitor
Amino Acid Sequence
Animals
Aprotinin
Base Sequence
Cattle
Chymases
Cloning, Molecular
DNA, Complementary
Humans
Mast Cells
Molecular Sequence Data
Phylogeny
Sequence Homology, Amino Acid
Serine Endopeptidases
Tryptases
Pallaoro, M., Gambacurta, A., Fiorucci, L., Mignogna, G., Barra, D., Ascoli, F. (1996). 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. EUROPEAN JOURNAL OF BIOCHEMISTRY, 237(1), 100-105 [10.1111/j.1432-1033.1996.0100t.x].
Pallaoro, M; Gambacurta, A; Fiorucci, L; Mignogna, G; Barra, D; Ascoli, F
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/258262
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