Abstract
Incubation of rat hepatocytes with glucose results in a decrease in the amount of glycogen synthase activity found in supernatants obtained after centrifugation of cell homogenates at 9200 g. The enzymic activity was quantitatively recovered in the sediments. This effect of translocation was dose- and time-dependent and correlated with the amount of immunoreactive enzyme determined by immunoblotting in both fractions. Hydrolysis by alpha-amylase of glycogen accumulated upon incubation with the sugar did not affect the translocation pattern. Translocation was also observed when cells were incubated with 2-deoxyglucose, which did not result in accumulation of glycogen. Immunocytochemical evidence indicates that glucose induces the aggregation of glycogen synthase molecules into clusters which are recovered in the sediments. These results indicate that glucose, in addition to activating glycogen synthase, may trigger changes in the localization of the enzyme in the cell.
Full text
PDF





Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ariño J., Mor A., Bosch F., Baanante I. V., Guinovart J. J. Hormonal effects on the phosphorylation of glycogen synthase in rat hepatocytes. FEBS Lett. 1984 May 21;170(2):310–314. doi: 10.1016/0014-5793(84)81334-4. [DOI] [PubMed] [Google Scholar]
- Ballard F. J., Oliver I. T. The effect of concentration on glucose phosphorylation and incorporation into glycogen in the livers of foetal and adult rats and sheep. Biochem J. 1964 Jul;92(1):131–136. doi: 10.1042/bj0920131. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barber A. A., Orrell S. A., Jr, Bueding E. Association of enzymes with rat liver glycogen isolated by rate-zonal centrifugation. J Biol Chem. 1967 Sep 25;242(18):4040–4044. [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
- Ciudad C. J., Massagué J., Guinovart J. J. The inactivation of glycogen phosphorylase is not a prerequisite for the activation of liver glycogen synthase. FEBS Lett. 1979 Mar 15;99(2):321–324. doi: 10.1016/0014-5793(79)80982-5. [DOI] [PubMed] [Google Scholar]
- Ciudad C., Camici M., Ahmad Z., Wang Y., DePaoli-Roach A. A., Roach P. J. Control of glycogen synthase phosphorylation in isolated rat hepatocytes by epinephrine, vasopressin and glucagon. Eur J Biochem. 1984 Aug 1;142(3):511–520. doi: 10.1111/j.1432-1033.1984.tb08315.x. [DOI] [PubMed] [Google Scholar]
- Czech M. P. Cellular basis of insulin insensitivity in large rat adipocytes. J Clin Invest. 1976 Jun;57(6):1523–1532. doi: 10.1172/JCI108422. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Wulf H., Hers H. G. The stimulation of glycogen synthesis and of glycogen synthetase in the liver by the administration of glucose. Eur J Biochem. 1967 Jul;2(1):50–56. doi: 10.1111/j.1432-1033.1967.tb00104.x. [DOI] [PubMed] [Google Scholar]
- De Wulf H., Stalmans W., Hers H. G. The effect of glucose and of a treatment by glucocorticoids on the activation in vitro of liver glycogen synthetase. Eur J Biochem. 1970 Jul;15(1):1–8. doi: 10.1111/j.1432-1033.1970.tb00967.x. [DOI] [PubMed] [Google Scholar]
- GORNALL A. G., BARDAWILL C. J., DAVID M. M. Determination of serum proteins by means of the biuret reaction. J Biol Chem. 1949 Feb;177(2):751–766. [PubMed] [Google Scholar]
- Griffiths G., McDowall A., Back R., Dubochet J. On the preparation of cryosections for immunocytochemistry. J Ultrastruct Res. 1984 Oct;89(1):65–78. doi: 10.1016/s0022-5320(84)80024-6. [DOI] [PubMed] [Google Scholar]
- Katz J., Golden S., Wals P. A. Glycogen synthesis by rat hepatocytes. Biochem J. 1979 May 15;180(2):389–402. doi: 10.1042/bj1800389. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lawrence J. C., Jr, Larner J. Activation of glycogen synthase in rat adipocytes by insulin and glucose involves increased glucose transport and phosphorylation. J Biol Chem. 1978 Apr 10;253(7):2104–2113. [PubMed] [Google Scholar]
- Maddaiah V. T., Madsen N. B. Studies on the biological control of glycogen metabolism in liver. 3. Subcellular distribution of glycogen metabolizing enzymes. Can J Biochem. 1968 Jun;46(6):521–525. doi: 10.1139/o68-081. [DOI] [PubMed] [Google Scholar]
- Massagué J., Guinovart J. J. Insulin control of rat hepatocyte glycogen synthase and phosphorylase in the absence of glucose. FEBS Lett. 1977 Oct 15;82(2):317–320. doi: 10.1016/0014-5793(77)80610-8. [DOI] [PubMed] [Google Scholar]
- Masters C. Interactions between glycolytic enzymes and components of the cytomatrix. J Cell Biol. 1984 Jul;99(1 Pt 2):222s–225s. doi: 10.1083/jcb.99.1.222s. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Méjean C., Pons F., Benyamin Y., Roustan C. Antigenic probes locate binding sites for the glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase, aldolase and phosphofructokinase on the actin monomer in microfilaments. Biochem J. 1989 Dec 15;264(3):671–677. doi: 10.1042/bj2640671. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stalmans W., De Wulf H., Hue L., Hers H. G. The sequential inactivation of glycogen phosphorylase and activation of glycogen synthetase in liver after the administration of glucose to mice and rats. The mechanism of the hepatic threshold to glucose. Eur J Biochem. 1974 Jan 3;41(1):127–134. doi: 10.1111/j.1432-1033.1974.tb03252.x. [DOI] [PubMed] [Google Scholar]
- Tan A. W. A simplified method for the preparation of pure UDP[14C] glucose. Biochim Biophys Acta. 1979 Feb 1;582(3):543–547. doi: 10.1016/0304-4165(79)90146-6. [DOI] [PubMed] [Google Scholar]
- Tan A. W., Nuttall F. Q. In vivo phosphorylation of liver glycogen synthase. Isolation of the 32P-labeled enzyme and studies on the nature of the bound [32P]phosphates. J Biol Chem. 1985 Apr 25;260(8):4751–4757. [PubMed] [Google Scholar]
- Thomas J. A., Schlender K. K., Larner J. A rapid filter paper assay for UDPglucose-glycogen glucosyltransferase, including an improved biosynthesis of UDP-14C-glucose. Anal Biochem. 1968 Oct 24;25(1):486–499. doi: 10.1016/0003-2697(68)90127-9. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]




