IDENTIFICATION OF TARGETING PROTEINASE FOR RAT ALPHA(1)-MACROGLOBULININ-VIVO - MAST-CELL TRYPTASE IS A MAJOR COMPONENT OF THE ALPHA(1)-MACROGLOBULIN-PROTEINASE COMPLEX ENDOCYTOSED INTO RAT-LIVER LYSOSOMES
A. Tsuji et al., IDENTIFICATION OF TARGETING PROTEINASE FOR RAT ALPHA(1)-MACROGLOBULININ-VIVO - MAST-CELL TRYPTASE IS A MAJOR COMPONENT OF THE ALPHA(1)-MACROGLOBULIN-PROTEINASE COMPLEX ENDOCYTOSED INTO RAT-LIVER LYSOSOMES, Biochemical journal, 298, 1994, pp. 79-85
The alpha(1)-macroglobulin-proteinase complex endocytosed into rat liv
er lysosomes was purified by a series of column chromatographic steps
on concanavalin A-Sepharose, Sephacryl S-300, DEAE-cellulose and TSK g
el DEAE-5PW columns. The complex contained no detectable alpha(2)-macr
oglobulin. Studies on the substrate specificity indicated that the com
plex had tryptase-like activities towards various synthetic substrates
, but no elastase, chymotrypsin, cathepsin-B and cathepsin-L activitie
s. The proteinase activity was completely inhibited by di-isopropyl fl
uorophosphate, leupeptin and antipain, indicating that the proteinase
bound to alpha(1)-macroglobulin is a serine proteinase. Two protein ba
nds (62 and 59 kDa) of the complex were labelled with [H-3]di-isopropy
l fluorophosphate and both bands cross-reacted with anti-(mast-cell tr
yptase)antibody. These results suggest that mast-cell tryptase is a ma
jor targeting proteinase for alpha 1(-)macroglobulin in vivo. The main
alpha-macroglobulin-proteinase complex in the adjuvant-treated rats w
as also the alpha(1)-macroglobulin-tryptase complex, even though the p
lasma level of alpha(2)-macroglobulin was elevated.