Investigation of glycosylation processes in mitochondria and microsomal membranes from human skeletal muscle.
Glycoconjugates are directly involved in major skeletal muscle functions. As little is known about glycosylation processes in muscle, we investigated glycoconjugate synthesis in subcellular fractions from human skeletal muscle tissue. Mitochondria and microsomal membranes were prepared from muscle biopsies by thorough mechanical disruption and differential centrifugations. This procedure resulted in the isolation of intact mitochondria (1 mg protein/g muscle) and of a microsomal fraction (1.5 mg protein/g muscle). Glycosyltransferases were studied in both subcellular fractions using either dolichylmonophosphate as a polyprenic acceptor or chemically modified fetuin as a glycoprotein substrate. Our results provide evidence for high rates of glycosylation in muscle. The highest activities were obtained with GDP-mannose: dilichylmonophosphate mannosyltransferase, a key enzyme in glycosylation process (220 pmol/mg per h in mitochondria and 1,550 pmol/mg per h in microsomal membranes). Substantial individual variations were observed for dolichol pathway glycosyltransferases but low individual variations were found for glycosyltransferases involved in maturation of glycoproteins. The role which glycosylation defects may play in muscle dysfunction has yet to be defined.
Gasnier
F
F
University of Lyon, Lyon-Sud Medical School, Department of Biochemistry, INSERM U. 189-CNRS, Oullins France.
Lerme
F
F
Rousson
R
R
Roussouly
P
P
Vaganay
E
E
Louisot
P
P
Gateau-Roesch
O
O
D000293:Adolescent; D000328:Adult; D004286:Dolichols / Q000378:metabolism; D005260:Female; D005690:Galactose / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D005964:Glucosyltransferases / Q000378:metabolism; D006801:Humans; D008297:Male; D008566:Membranes / Q000378:metabolism; D008861:Microsomes / Q000201:enzymology / Q000378:metabolism*; D008875:Middle Aged; D008931:Mitochondria, Muscle / Q000201:enzymology / Q000378:metabolism*; D009124:Muscle Proteins / Q000378:metabolism; D009132:Muscles / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D009135:Muscular Diseases / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000378:metabolism
D004286:Dolichols; D009124:Muscle Proteins; D000509:alpha-Fetoproteins; D005964:Glucosyltransferases; D005947:Glucose; D005690:Galactose