Glucose uptake and metabolism by cultured human skeletal muscle cells: Rate-limiting steps

Laureta M. Perriott, Tetsuro Kono, Richard R. Whitesell, Susan M. Knobel, David W. Piston, Daryl K. Granner, Alvin C. Powers, James M. May

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

To use primary cultures of human skeletal muscle cells to establish defects in glucose metabolism that underlie clinical insulin resistance, it is necessary to define the rate-determining steps in glucose metabolism and to improve the insulin response attained in previous studies. We modified experimental conditions to achieve an insulin effect on 3-O-methylglucose transport that was more than twofold over basal. Glucose phosphorylation by hexokinase limits glucose metabolism in these cells, because the apparent Michaelis-Menten constant of coupled glucose transport and phosphorylation is intermediate between that of transport and that of the hexokinase and because rates of 2-deoxyglucose uptake and phosphorylation are less than those of glucose. The latter reflects a preference of hexokinase for glucose over 2-deoxyglucose. Cellular NAD(P)H autofluorescence, measured using two-photon excitation microscopy, is both sensitive to insulin and indicative of additional distal control steps in glucose metabolism. Whereas the predominant effect of insulin in human skeletal muscle cells is to enhance glucose transport, phosphorylation, and steps beyond, it also determines the overall rate of glucose metabolism.

Original languageEnglish
Pages (from-to)E72-E80
JournalAmerican Journal of Physiology - Endocrinology and Metabolism
Volume281
Issue number1 44-1
StatePublished - Oct 10 2001
Externally publishedYes

Keywords

  • Glucose transport
  • Hexokinase
  • Rate-determining step
  • Substrate specificity
  • Transport-metabolism coupling

Fingerprint Dive into the research topics of 'Glucose uptake and metabolism by cultured human skeletal muscle cells: Rate-limiting steps'. Together they form a unique fingerprint.

  • Cite this

    Perriott, L. M., Kono, T., Whitesell, R. R., Knobel, S. M., Piston, D. W., Granner, D. K., Powers, A. C., & May, J. M. (2001). Glucose uptake and metabolism by cultured human skeletal muscle cells: Rate-limiting steps. American Journal of Physiology - Endocrinology and Metabolism, 281(1 44-1), E72-E80.