Low density lipoprotein degradation by secretory granules of rat mast cells. Sequential degradation of apolipoprotein B by granule chymase and carboxypeptidase A.
The secretory granules of rat serosal mast cells are able efficiently to degrade the apolipoprotein B component of low density lipoproteins (LDL) Kokkonen, J. O., and Kovanen, P. T. (1985) J. Biol. Chem. 260, 14756-14763). The granules are known to contain two neutral proteases with complementary specificities: a chymotrypsin-like endopeptidase called chymase, and an exopeptidase, the granule carboxypeptidase A. The role of this enzyme pair in the proteolytic degradation of LDL was studied with the aid of specific enzyme inhibitors. Incubation of LDL with intact granules (both enzymes active) led to the formation of numerous low molecular weight peptides and the liberation of free amino acids, most of which (95%) were aromatic (Phe, Tyr, Trp) or branched-chain aliphatic (Leu, Ile, Val). Selective inhibition of granule carboxypeptidase A (leaving chymase active) blocked the liberation of free amino acids, but left the formation of peptides uninhibited. On the other hand, selective inhibition of granule chymase (leaving carboxypeptidase A active) totally abolished the proteolytic degradation of LDL. The results are consistent with a model according to which the proteolytic degradation of LDL by mast cell granules results from coordinated action of the two granule-bound enzymes, whereby the chymase first cleaves peptides from the apolipoprotein B of LDL, and thereafter the carboxypeptidase A cleaves amino acids from the peptides formed.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001055:Apolipoproteins B / Q000378:metabolism*; D002268:Carboxypeptidases / Q000032:analysis / Q000502:physiology*; D043422:Carboxypeptidases A; D053818:Chymases; D003594:Cytoplasmic Granules / Q000201:enzymology / Q000378:metabolism*; D010450:Endopeptidases / Q000032:analysis / Q000502:physiology*; D066298:In Vitro Techniques; D008077:Lipoproteins, LDL / Q000032:analysis / Q000378:metabolism*; D008297:Male; D008407:Mast Cells / Q000378:metabolism*; D010455:Peptides / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D012697:Serine Endopeptidases
D000596:Amino Acids; D001055:Apolipoproteins B; D008077:Lipoproteins, LDL; D010455:Peptides; D002268:Carboxypeptidases; D010450:Endopeptidases; D043422:Carboxypeptidases A; D012697:Serine Endopeptidases; D053818:Chymases