Secretory granule proteases in rat mast cells. Cloning of 10 different serine proteases and a carboxypeptidase A from various rat mast cell populations.

"the processed mature protease of 227 aa has a molecular mass of 25193 (excluding potential carbohydrate additions) a net charge of +19 1 (arg + lys = 34; asp + glu = 16) and contains no potential n -glycosylation sites (the nucleotide sequence is deposited in genbank under the accession number u67915) a nearly full-length cdna clone for the rat chymase rmcp-5 was isolated (figs2 a and 5 a ).; the processed proteolytically active rmcp-5 is a basic protein with a net charge of +14 1 (arg + lys = 29; asp + glu = 16) (the nucleotide sequence is deposited in genbank under the accession number u67908) the sequence differs in two positions compared with the previously published sequence of rmcp-5 (two additional nucleotides t in pos810 and a g in pos811) ( ).; the processed proteolytically active rmcp-6 is an acidic protein with a net charge of -4 6 (arg + lys = 19; asp + glu = 25) (the nucleotide sequence is deposited in genbank under the accession number u67909) the sequence differs in five positions from the previously published sequence of the rat tryptase (a pos399 c pos432 c pos748 t pos946 and one base pair deleted at position 1024) ( ).; the processed proteolytically active rmcp-7 is an acidic protein with a net charge of -4 6 (arg + lys = 17; asp + glu = 23) (the nucleotide sequence is deposited in genbank under the accession number u67910) a nearly full-length cdna clone for rat mc carboxypeptidase a (r-cpa) was isolated (figs4 and 5 d ).; the processed proteolytically active r-cpa is a basic protein with a net charge of +18 1 (arg + lys = 46; asp + glu = 29) (the nucleotide sequence is deposited in genbank under the accession number u67914) mucosal mcs are difficult to isolate as a pure population.; the processed proteolytically active rmcp-3 is a basic protein with a net charge of +9 2 (arg + lys = 28; asp + glu = 20) (the nucleotide sequence is deposited in genbank under the accession number u67888) a cdna clone containing the entire coding region for rmcp-4 was isolated.; the mature basic protein has a net charge of +12 1 (arg + lys = 26; asp + glu = 15) (the nucleotide sequence is deposited in genbank under the accession number u67907) a cdna clone containing the entire coding region for rmcp-8 was isolated (figs2 b and 5 a ).; the active protease is a basic protein with a net charge of +10 6 (arg + lys = 27; asp + glu = 17) (the nucleotide sequence is deposited in genbank under the accession number u67911) a nearly full-length cdna clone for rmcp-9 was isolated (figs2 b and 5 b ).; the active basic enzyme has228 aa with a molecular mass of 25249 a net charge of +12 3 (arg + lys = 27; asp + glu = 15) and two putative glycosylation sites at positions132 and 160 (the nucleotide sequence is deposited in genbank under the accession number u67912) a clone containing the partial sequence of rmcp-10 was isolated by pcr amplification with the his-ser primers (see above).; the obtained sequence covers the central region between the his and ser residues of the catalytic triad (figs2 b and 5 b ) (the nucleotide sequence is deposited in genbank under the accession number u67913) except for a single base pair difference this sequence was found to be identical to a cdna previously isolated from rat duodenum by amerik et al ( ) (granzyme-like protein ii)."

"We are grateful to Dr. Anders Karlström for performing NH2-terminal sequence analysis. We would also like to thank Dr. David Eaker for linguistic revision of the manuscript. This investigation was supported by grants from the Swedish Natural Sciences Research Council."