Low levels of glucose-6-phosphate hydrolysis in the sarcoplasmic reticulum of skeletal muscle: involvement of glucose-6-phosphatase.
Glucose-6-phosphate hydrolysis was measured in a fraction obtained from rabbit fast-twitch skeletal muscle and corresponding to total sarcoplasmic reticulum, as well as in three subfractions containing longitudinal tubules, terminal cisternae or both structures. In all cases the levels of hydrolysis measured both in native and disrupted membranes were approximately 60-100 times lower than the microsomal glucose-6-phosphatase activity of the corresponding livers. In contrast to liver microsomes, most (up to 80%) of the glucose-6-phosphate hydrolysing activity in muscle sarcoplasmic reticulum membranes was not inactivated by pH 5.0 pre-incubation indicating that it was not catalysed by the specific glucose-6-phosphatase enzyme. Osmotically induced changes in light-scattering intensity of sarcoplasmic reticulum vesicles revealed that, in contrast to liver microsomes, sarcoplasmic reticulum vesicles were not selectively permeable to glucose-6-phosphate as mannose-6-phosphate was also permeable and in addition they were poorly permeable to glucose. Immunoblot experiments using antibodies raised against the glucose-6-phosphatase enzyme, and liver endoplasmic reticulum glucose and Pi translocases, failed to detect the presence of these protein components in sarcoplasmic reticulum membranes. Southern blot analysis of reverse transcriptase-polymerase chain reaction products from rat muscle revealed that glucose-6-phosphatase mRNA is present in muscle. Quantification of Northern blot analysis of liver and muscle mRNA indicated that muscle contains less than 2% of the amount of glucose-6-phosphate mRNA found in corresponding livers. We conclude that very low levels of specific glucose-6-phosphatase (e.g. as in liver; E.C. 3.1.3.9) are present in muscle sarcoplasmic reticulum and that the muscle and liver glucose-6-phosphatase systems have several different properties.
Gamberucci
A
A
Instituto di Patologia Generale, Università di Siena, Italy.
Marcolongo
P
P
Fulceri
R
R
Giunti
R
R
Watkins
S L
SL
Waddell
I D
ID
Burchell
A
A
Benedetti
A
A
D000818:Animals; D015139:Blotting, Southern; D015342:DNA Probes; D018076:DNA, Complementary; D005952:Glucose-6-Phosphatase / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D019298:Glucose-6-Phosphate / Q000378:metabolism*; D006868:Hydrolysis; D015151:Immunoblotting; D008360:Mannosephosphates / Q000378:metabolism; D008862:Microsomes, Liver / Q000201:enzymology; D018656:Muscle Fibers, Fast-Twitch / Q000378:metabolism; D018482:Muscle, Skeletal / Q000737:chemistry / Q000378:metabolism; D010539:Permeability; D016133:Polymerase Chain Reaction; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*
D015342:DNA Probes; D018076:DNA, Complementary; D008360:Mannosephosphates; C027693:mannose-6-phosphate; D019298:Glucose-6-Phosphate; D005952:Glucose-6-Phosphatase