Characterization of collagen hydroxylysyl glycosyltransferases as mainly intramembranous microsomal enzymes.
The localization of collagen hydroxylysine galactosyl- and galactosyl-hydroxylysine glucosyltransferases in purified chick embryo bone microsomes was studied by differential solubilization with nonionic detergents. Brij-35 (polyoxyethylene 25-lauryl ether) which selectively releases intracisternal proteins, and Triton X-100, whose specificity varies with its concentration, were used in the presence or absence of high ionic strength NaCl. These methods were used previously to characterize prolyl hydroxylase as intracisternal and lysyl hydroxylase as mainly intramembranous. The distribution of both glycosyltransferases within microsomes was similar to that of lysyl hydroxylase; approximately 70-80% of their activities are intramembranous with the remainder intracisternal. Collagen hydroxylysine glucosyltransferase differed from prolyl and lysyl hydroxylase and the galactosyltransferase in that its activity in vitro was apparently inhibited by membrane vesicles, even in the presence of detergents at concentrations which permeabilize the membrane. Accurate measurement of its activity could be achieved only by its separation from vesicles after detergent treatment. The common location of the major portion of lysyl hydroxylase and the glycosyltransferase activities suggests that they may act as a multienzyme complex to preferentially modify certain lysyl residues in nascent procollagen chains as they traverse the membrane of the endoplasmic reticulum. Since these enzymes do not act on helical collagen, their physical separation from prolyl hydroxylase may ensure that modifications of lysine residues occur prior to formation of hydroxyproline, which stabilizes the helical form.
D000818:Animals; D001842:Bone and Bones / Q000201:enzymology*; D002642:Chick Embryo; D005700:Galactosyltransferases / Q000378:metabolism*; D005964:Glucosyltransferases / Q000378:metabolism*; D007425:Intracellular Membranes / Q000201:enzymology; D008861:Microsomes / Q000201:enzymology*; D017830:Octoxynol; D000077423:Polidocanol; D011092:Polyethylene Glycols; D012995:Solubility
D000077423:Polidocanol; D011092:Polyethylene Glycols; D017830:Octoxynol; D005700:Galactosyltransferases; D005964:Glucosyltransferases; C023236:UDPgalactose-collagen galactosyltransferase; C023247:UDP glucose-collagen glucosyltransferase