Identification and molecular cloning of a novel mouse mucosal mast cell serine protease

• Published in: The Journal of biological chemistry Vol. 265; no. 1; pp. 423 - 429
• Main Authors: SERAFIN, W. E, REYNOLDS, D. S, ROGELJ, S, LANE, W. S, CONDER, G. A, JOHNSON, S. S, AUSTEN, K. F, STEVENS, R. L
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 05.01.1990
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Animals; Base Sequence; Biological and medical sciences; Biotechnology; Cell Line, Transformed; Cell Transformation, Viral; Cloning, Molecular; Cytoplasmic Granules - enzymology; DNA - genetics; DNA - isolation & purification; DNA Probes; Enzymes and enzyme inhibitors; Female; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Hydrolases; Kirsten murine sarcoma virus; mast cells; Mast Cells - enzymology; Methods. Procedures. Technologies; Mice; Mice, Inbred BALB C; Molecular cloning; Molecular Sequence Data; Molecular Weight; Nucleic Acid Hybridization; RNA - genetics; Serine Endopeptidases - analysis; Serine Endopeptidases - genetics; Nucleotide sequence; Proteinase; Rodentia; Identification; Serine enzyme; Vertebrata; Mammalia; Cell line; Mouse; Blotting assay; Mast cell; Molecular cloning; Aminoacid sequence
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/s0021-9258(19)40247-0

A novel 28,000 Mr serine protease, designated mouse mast cell protease-2 (MMCP-2), that is stored in the secretory granules of Kirsten sarcoma virus-immortalized mouse mast cells (KiSV-MC) has been identified and its NH2-terminal amino acid sequence has been determined. Analysis of a 953-base pair cDNA that encodes MMCP-2 revealed that this serine protease is a basically charged protein, possessing the histidine-aspartic acid-serine charge relay system that is characteristic of other serine proteases. DNA blot analysis using the full-length MMCP-2 cDNA indicated the existence of a family of highly related serine protease genes in the mouse genome. When the same DNA blot was probed with the 149-base pair KpnI---3' fragment of the cDNA, the probe hybridized to a single DNA fragment, thereby demonstrating that this 3' fragment could be used as a gene-specific probe. The presence of high levels of the MMCP-2 mRNA transcript in the intestines of nematode-infected mice, and its absence in mouse bone marrow-derived mast cells and peritoneal cavity-derived connective tissue mast cells, suggest that this member of the mouse mast cell protease family is preferentially expressed late in the differentiation of mucosal mast cells.