Skip to main content

Binding and molecular weight properties of the insulin receptor from omental and subcutaneous adipocytes in human obesity

Summary

The insulin binding properties and the molecular weights of the insulin receptor and its insulin binding subunit were studied in omental and subcutaneous adipocytes prepared from obese- and normal-weight subjects. Insulin binding by such adipocytes was decreased in obesity when the binding activity was expressed per unit of cell surface area. No significant difference from the lean controls was evident, however, when binding was calculated on a per cell basis, indicating that the total receptor content of the cells from the obese subjects was not altered. In addition, the normal difference in the receptor binding affinities previously reported between omental and subcutaneous cells from lean individuals was unaffected by the obese condition. Studies of the molecular weight of the non-reduced insulin receptor in fat cell membranes prepared from pieces of omental and subcutaneous fat demonstrated a major receptor species of 390–425K Mr. In contrast, adipocytes isolated by collagenase treatment of the fat had heterogenous non-reduced receptor species of Mr 355K, 285K and small amounts of 427K and 182K. Although different non-reduced receptor species were evident depending on the adipocyte receptor preparation (e.g. isolated adipocytes or fat cell membranes), no differences were found between obese and lean controls or between subcutaneous and omental receptors when the appropriate comparisons were made. Upon sulphydryl reduction, all receptor preparations had a major binding subunit of 125K Mr. In conclusion, obesity is characterized by a dilution of the insulin receptor over the adipocyte cell surface in the absence of a change in total cellular content of receptors. The difference in insulin binding affinities between omental and subcutaneous adipocytes could not be explained by an alteration in receptor molecular weight.

References

  1. Kreisberg R, Boshell B, DiPlacido J, Roddman R (1967) Insulin secretion in obesity. N Engl J Med 276: 314–319

    Google Scholar 

  2. Olefsky J (1981) Insulin resistance and insulin action. Diabetes 30: 148–162

    Google Scholar 

  3. Olefsky J (1976) Decreased insulin binding to adipocytes and circulating monocytes in obesity. J Clin Invest 57: 1165–1172

    Google Scholar 

  4. Kolterman O, Reaven G, Olefsky J (1979) Relationship between in vivo insulin resistance and decreased insulin receptors in obese man. J Clin Endocrinol Metab 48: 487–494

    Google Scholar 

  5. Harrison L, Martin F, Melick R (1976) Correlation between insulin receptor binding in isolated fat cells and insulin sensitivity in obese human subjects. J Clin Invest 58: 1435–1441

    Google Scholar 

  6. Kolterman O, Insel J, Saekow M, and Olefsky J (1980) Mechanisms of insulin resistance in human obesity. J Clin Invest 65: 1272–1284

    Google Scholar 

  7. Amatruda M, Livingston J, Lockwood D (1975) Insulin receptor: role in the resistance of human obesity to insulin. Science 188: 264–266

    Google Scholar 

  8. Pedersen O, Hijollunk E, Schwartz N (1982) Insulin receptor binding and insulin action in human fat cells: effects of obesity and fasting. Metabolism 31: 884–895

    Google Scholar 

  9. Lönnroth P, DiGirolamo M, Krotkiewski M, Smith U (1983) Insulin binding and responsiveness in fat cells from patients with reduced glucose tolerance and type II diabetes. Diabetes 32: 748–754

    Google Scholar 

  10. Borkan G, Cerzof S, Robbins A, Hults D, Silbert C, Silbert J (1982) Assessment of abdominal fat content by computed tomography. Am J Clin Nutr 36: 172–177

    Google Scholar 

  11. Bolinder J, Kager L, Östman J, Arner P (1983) Differences at the receptor and postreceptor levels between human omental and subcutaneous adipose tissue in the action of insulin on lipolysis. Diabetes 32: 117–123

    Google Scholar 

  12. Sjöstrom L, Björntorp P, Vrana J (1971) Microscopic fat cells: size measurements on frozen adipose tissue in comparison with automatic determinations of osimum-fixed fat cells. J Lipid Res 12: 521–530

    Google Scholar 

  13. Hirsch J, Gallian E (1968) Method for the determinations of adipose cell size and cell number in man and animals. J Lipid Res 9: 110–119

    Google Scholar 

  14. Zinder Z, Shapiro B (1971) Effect of cell size on epinephrine- and ACTH-induced fatty acid release from isolated fat cells. J Lipid Res 12: 91–95

    Google Scholar 

  15. Rodbell M (1964) Metabolism of isolated fat cells. I. Effects of hormone on glucose metabolism and lipolysis. J Biol Chem 239: 375–380

    Google Scholar 

  16. Belsham G, Denton R, Tanner M (1980) Use of novel rapid preparation of fat-cell plasma membranes employing Percoll to investigate the effects of insulin and adrenaline on membrane protein phosphorylation within intact fat-cells. Biochem J 192: 457–467

    Google Scholar 

  17. Gliemann J, Sonne O (1978) Binding and receptor-mediated degradation of insulin in adipocytes. J Biol Chem 253: 7857–7863

    Google Scholar 

  18. Scatchard G (1949) The attraction of protein for small molecules and ions. Ann NY Acad Sci 51: 660–672

    Google Scholar 

  19. Pilch P, Czech M (1979) Interaction of cross-linking agents with the insulin effector system of isolated fat cells. J Biol Chem 254: 3375–3381

    Google Scholar 

  20. Weber K, Pringle J, Osborn M (1972) Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol 26: 3–27

    Google Scholar 

  21. Laemmli U (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    Google Scholar 

  22. Livingston J, Purvis B (1980) Effects of wheat germ agglutinin on insulin binding and insulin sensitivity of fat cells. Am J Physiol 238: E267-E275

    Google Scholar 

  23. Lerea K, Livingston J (1983) The different receptor species of liver have similar complex insulin binding properties. Biochem Biophys Res Commun 114: 1042–1047

    Google Scholar 

  24. Massague J, Pilch P, Czech M (1980) Electrophoretic resolution of three major insulin-receptor structures with unique subunit stoichiometries. Proc Natl Acad Sci (USA) 77: 7137–7141

    Google Scholar 

  25. Ciaraldi T, Kolterman O, Olefsky J (1981) Mechanism of the postreceptor defect in insulin action in human obesity. J Clin Invest 68: 875–880

    Google Scholar 

  26. Arner P, Bolinder J, Engfeldt P, Ostman J (1981) The antilipolytic effect of insulin in human adipose tissue in obesity, diabetes mellitus, hyperinsulinemia, and starvation. Metabolism 30: 753–760

    Google Scholar 

  27. Massague J, Pilch P, Czech M (1981) A unique proteolytic cleavage site on the β subunit of the insulin receptor. J Biol Chem 256: 3182–3190

    Google Scholar 

  28. Berhanu P, Kolterman O, Baron A, Tsai P, Olefsky J, Brandenburg D (1983) Insulin receptors in isolated human adipocytes. Characterization by photoaffinity labeling and evidence for internalization and cellular processing. J Clin Invest 72: 1958–1970

    Google Scholar 

  29. Rodbell, M (1964) Localization of lipoprotein lipase in fat cells of rat adipose tissue. J Biol Chem 239: 753–755

    Google Scholar 

  30. Mott D, Howard B, Bennett P (1979) Stoichiometric binding and regulation of insulin receptors on human diploid fibroblasts using physiologic insulin levels. J Biol Chem 254: 8762–8767

    Google Scholar 

  31. Gould R, Ginsberg B, Spector A (1982) Lipid effects on the binding properties of a reconstituted insulin receptor. J Biol Chem 257: 477–484

    Google Scholar 

  32. Mature J, Hollenberg M (1978) Insulin receptor: interaction with non-receptor glycoprotein from liver cell membranes. Proc Natl Acad Sci (USA) 75: 3070–3074

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Livingston, J.N., Lerea, K.M., Bolinder, J. et al. Binding and molecular weight properties of the insulin receptor from omental and subcutaneous adipocytes in human obesity. Diabetologia 27, 447–453 (1984). https://doi.org/10.1007/BF00273909

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00273909

Key words

  • Insulin receptor
  • obesity
  • adipocyte
  • subcutaneous fat
  • omental fat