Differential inhibition of rat mast cell proteinase I and II by members of the alpha-1-proteinase inhibitor family of serine proteinase inhibitors.
Rat mast cell proteinase II (RMCP II) from mucosal mast cells was titrated into rat serum, and the resulting serine proteinase inhibitor (serpin)-enzyme complex was purified by affinity chromatography on anti-RMCP II-Sepharose 4B and by Mono-Q anion-exchange. The purified complex was used to raise polyclonal antibodies which, after cross-absorption against RMCP II-Sepharose 4B, were specific for serpin and were used to affinity purify two rat serpin molecules (RSI and RSII) that inhibit RMCP II in rat serum. The kinetic constants characterizing the interaction between RMCP II and RSI and RSII are ka, 2.2 x 10(5) and 1.65 x 10(5) M-1 s-1, respectively; Ki, 3.6 x 10(-10) and 1.0 x 10(-9) M; and kd, 7.9 x 10(-5) and 1.65 x 10(-4) s-1. Amino-terminal sequence analysis indicated that RSI and RSII are distinct, differing at the amino-terminal residues, and are products of the rat Spi-1 locus. Rat mast cell proteinase I (RMCP I) from connective tissue mast cells cleaved both RSI and RSII and was not inhibited.
Pirie-Shepherd
S R
SR
Moredun Research Institute, Edinburgh, Scotland.
Miller
H R
HR
Ryle
A
A
D000595:Amino Acid Sequence; D000818:Animals; D002852:Chromatography, Ion Exchange; D053818:Chymases; D008297:Male; D008407:Mast Cells / Q000201:enzymology*; D008969:Molecular Sequence Data; D051381:Rats; D011919:Rats, Inbred Strains; D012697:Serine Endopeptidases / Q000302:isolation & purification / Q000378:metabolism; D015842:Serine Proteinase Inhibitors; D015843:Serpins / Q000502:physiology*
D015842:Serine Proteinase Inhibitors; D015843:Serpins; D012697:Serine Endopeptidases; C057818:cytotoxic T lymphocyte-specific serine protease; D053818:Chymases