Summary
The human insulin receptor exists in two isoforms (HIR-A α-subunit 719 amino acids and HIR-B α-subunit 731 amino acids) which are generated by alternative splicing of a small exon and display distinct patterns of tissue-specific expression. Using the polymerase chain reaction we have recently shown that skeletal muscle of non-diabetic individuals contains predominantly mRNA encoding HIR-A while in skeletal muscle derived from subjects with Type 2 (non-insulin-dependent) diabetes mellitus similar amounts of each mRNA are expressed. We used a polyclonal antibody which discriminates between HIR-A and HIR-B to assess the isoform expression at the protein level. The antibody showed clearly distinct displacement of insulin binding in skeletal muscle membranes of non-diabetic subjects compared to Type 2 diabetic subjects (displacement of specific 125I-insulin binding: 13 non-diabetic subjects 70.0%±14.34, 12 Type 2 diabetic subjects 32.6%±17.45). A control antibody which does not discriminate between both isoforms showed similar displacement of 125I-insulin in membranes of non-diabetic and Type 2 diabetic subjects. These data suggest that the altered expression of receptor isotype mRNA in the skeletal muscle of Type 2 diabetic subjects leads to an altered receptor isoform pattern in the plasma membrane. While skeletal muscle membranes of non-diabetic subjects contain predominantly HIR-A, membranes of Type 2 diabetic subjects show an increased level of HIR-B in addition to HIR-A.
Article PDF
Similar content being viewed by others
References
Ullrich A, Bell JB, Chen EY et al. (1985) Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. Nature 313: 756–761
Ebina Y, Ellis L, Jarmagin K et al. (1985) The human insulin receptor c-DNA: the structural basis for hormone-activated transmembrane signalling. Cell 46: 747–758
Moller DE, Yokota A, Caro JF, Flier JS (1989) Tissue-specific expression of two alternatively spliced insulin receptor mRNAs in man. Mol Endocrinology 3: 1263–1269
Seino S, Bell GI (1989) Alternative splicing of human insulin receptor messenger RNA. Biochem Biophys Res Commun 159: 312–316
Mosthaf L, Grako D, Dull TJ et al. (1990) Functionally distinct insulin receptors generated by tissue-specific alternate splicing. EMBOJ 9: 2409–2413
Kellerer M, Lammers R, Ermel B et al. (1992) Different α-subunit structures of the human insulin receptors type A and B affect the tyrosine kinase activity of the β-subunit. Biochemistry 19: 4588–4596
Vogt B, Carrascosa J, Ermel B et al. (1991) The two isotypes of the human insulin receptor (HIR-A and HIR-B) follow different internalization kinetics. Biochem Biophys Res Commun 177: 1013–1018
McClain D (1991) Different ligand affinities of the two human insulin receptor splice variants are reflected in parallel changes in sensitivity for insulin action. Mol Endo 5: 734–739
Yamaguchi Y, Flier JS, Yokota A et al. (1991) Functional properties of two naturally occurring isoforms of the human insulin receptor in Chinese hamster ovary cells. Endocrinology 129: 2058–2066
Mosthaf L, Vogt B, Häring HU, Ullrich A (1991) Altered expression of insulin receptor types A and B in the skeletal muscle of non-insulin-dependent diabetes mellitus subjects. Proc Natl Acad Sci USA 88: 4728–4730
Benecke H, Flier JS, Moller DE (1992) Alternatively spliced variants of the insulin receptor protein. Expression in normal and diabetic human tissues. J Clin Invest 89: 2066–2070
Hansen T, Bjorback C, Vestergaard H, Bak JF, Pedersen O (1992) Alternatively spliced variants of the insulin receptor and its functional correlates in muscle from patients with type 2 diabetes and normal subjects. Diabetologia 35: A76
Mosthaf L, Eriksson J, Häring HU et al. (1993) Insulin receptor isotype expression correlates with risk of non-insulin-dependent diabetes. Proc Natl Acad Sci USA 90: in press
Sesti G, Marini MA, Montemurro A et al. (1992) The two naturally occurring human insulin receptor forms are immunologically distinct. Diabetes 41: 6–11
Soos MA, Siddle K, Baron MD et al. (1986) Monoclonal antibodies reacting with multiple epitopes on the human insulin receptor. Biochem J 235: 199–208
Obermaier-Kusser B, White MF, Pongratz DE et al. (1989) A defective intramolecular autoactivation cascade may cause the reduced kinase activity of the skeletal muscle insulin receptor from subjects with non-insulin-dependent diabetes mellitus. J Biol Chem 264: 9497–9503
Vogt B, Mühlbacher C, Carrascosa J et al. (1992) Subcellular distribution of GLUT 4 in skeletal muscle of lean type 2 (non-insulin-dependent) diabetic subjects in the basal state. Diabetologia 35: 456–463
Mühlbacher C, Karnieli E, Schaff P et al. (1988) Phorbol esters imitate in rat fat-cells the full effect of insulin on glucose-carrier translocation, but not on 3-0-methylglucose-transport activity. Biochem J 249: 865–870
Obermaier-Kusser B, Mühlbacher Ch, Mushack J et al. (1988) Regulation of glucose carrier activity by AlCl3 and phospholipase C in fat cells. Biochem J 256: 515–520
Karnieli E, Zarnowski MJ, Hissin PJ et al. (1981) Preparation and characterization of plasma membrane fraction from isolated rat fat cells. J Biol Chem 256: 4772–4777
Avruch J, Wallach DEH (1971) Erythrocyte membrane adenosine triphosphate activities in subjects with endogenous depression and healthy subjects. Biochem Biophys Acta 233: 334–347
Sesti G, Marini MA, Tullio AN et al. (1991) Altered expression of the two naturally occurring human insulin receptor variants in isolated adipocytes of non-insulin-dependent diabetes mellitus subjects. Biochem Biophys Res Commun 181: 1419–1424
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kellerer, M., Sesti, G., Seffer, E. et al. Altered pattern of insulin receptor isotypes in skeletal muscle membranes of Type 2 (non-insulin-dependent) diabetic subjects. Diabetologia 36, 628–632 (1993). https://doi.org/10.1007/BF00404072
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00404072