Abstract
Triton X-100 and exogenous dolichol monophosphate have been used to investigate the nature of enzymes responsible for the transfer of mannose, glucose and N-acetylglucosamine phosphate from nucleotide donors to dolichol monophosphate in vesicles derived from rough and smooth endoplasmic reticulum and mitochondria. Mitochondria were shown to contain the highest specific activities of these enzymes. The responses of the glycosyltransferases to increasing concentrations of Triton X-100 and the effect on these responses of exogenous dolichol monophosphate suggest that the enzymes for mannose and glucose transfer are less hydrophobic, and therefore less intrinsic, in the membrane than the enzyme for N-acetylglucosamine phosphate transfer. In smooth vesicles the results are consistent with mannosyl- and glucosyl-transferases being located at both inner and outer faces of the membrane. In rough vesicles and in mitochondria mannosyl- and glucosyl-transferases were confirmed at the outer face. There is, however, only one site of N-acetylglucosamine phosphate transfer, this being more hydrophobically located in the membrane than the other sites of glycosyl transfer. Mitochondrial enzyme activity closely resembled that of rough endoplasmic reticulum in response to Triton X-100 and exogenous dolichol monophosphate, and is probably associated with the outer membrane.
MeSH Terms
Acetylglucosamine/metabolism
Animals
Dolichol Phosphates/metabolism,pharmacology
Endoplasmic Reticulum/drug effects,metabolism
Glucose/metabolism
In Vitro Techniques
Liver/drug effects,metabolism
Male
Mannose/metabolism
Mitochondria, Liver/drug effects,metabolism
Polyethylene Glycols/pharmacology
Polyisoprenyl Phosphate Monosaccharides/biosynthesis
Polyisoprenyl Phosphates/metabolism
Rats
Time Factors
Chemicals
Dolichol Phosphates
Polyisoprenyl Phosphate Monosaccharides
Polyisoprenyl Phosphates
Polyethylene Glycols
Glucose
Mannose
Acetylglucosamine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Coolbear T
Mookerjea S
Hemming F W
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21 references, click to expand
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