Protein Tyrosine Phosphatase Regulation in Fibroblasts from Patients with an Insulin Receptor Gene Mutation

Abstract

Tyrosine phosphorylation of the insulin receptor is the initial event following receptor binding to insulin, and it induces further tyrosine phosphorylation of various intracellular molecules. This signaling is countered by protein tyrosine phosphatases (PTPases), which reportedly are associated with insulin resistance that can be reduced by regulation of PTPases. Protein tyrosine phosphatase 1B (PTP1B) and leukocyte antigen–related PTPase (LAR) are the PTPases implicated most frequently in insulin resistance and diabetes mellitus. Here, we show that PTP1B and LAR are expressed in human fibroblasts, and we examine the regulation of PTPase activity in fibroblasts from patients with an insulin receptor gene mutation as an in vitro model of insulin resistance. Total PTPase activity was significantly lower in the cytosolic and membrane fractions of fibroblasts with mutations compared with controls (p<0.05). Insulin stimulation of fibroblasts with mutations resulted in a significantly smaller increase in PTP1B activity compared with stimulation of wild-type fibroblasts (p<0.05). This indicates that insulin receptor gene mutations blunt increases in PTPase activity in response to insulin, possibly via a negative feedback mechanism. Our data suggest that the PTPase activity in patients with insulin receptor gene mutation and severe insulin resistance may differ from that in ordinary type 2 diabetes.

Key words

protein tyrosine phosphatase 1B - leukocyte antigen-related PTPase - diabetes mellitus - insulin resistance

References

• 1 Goldstein BJ. Protein-tyrosine phosphatases and the regulation of insulin action.  J Cell Biochem. 1992;  48 33-42
• 2 Ahmad F, Azevedo JL, Cortright R, Dohm GL, Goldstein BJ. Alterations in skeletal muscle protein-tyrosine phosphatase activity and expression in insulin-resistant human obesity and diabetes.  J Clin Invest. 1997;  100 449-458
• 3 Ahmad F, Considine RV, Bauer TL, Ohannesian JP, Marco CC, Goldstein BJ. Improved sensitivity to insulin in obese subjects following weight loss is accompanied by reduced protein-tyrosine phosphatases in adipose tissue.  Metabolism. 1997;  46 1140-1145
• 4 Taira M, Taira M, Hashimoto N, Shimada F, Suzuki Y, Kanatsuka A, Nakamura F, Ebina Y, Tatibana M, Makino H, Yoshida S. Human diabetes associated with a deletion of the tyrosine kinase domain of the insulin receptor.  Science. 1989;  245 63-66
• 5 Shimada F, Taira M, Suzuki Y, Hashimoto N, Nozaki O, Taira M, Tatibana M, Ebina Y, Tawata M, Onaya T, Makino H, Yoshida S. Insulin-resistant diabetes associated with partial deletion of insulin-receptor gene.  Lancet. 1990;  335 1179-1181
• 6 Nozaki O, Suzuki Y, Shimada F, Hashimoto N, Taira M, Hatanaka Y, Notoya Y, Kanashiro O, Makino H, Yoshida S. A glycine-1008 to valine mutation in the insulin receptor in a woman with type A insulin resistance.  J Clin Endocrinol Metab. 1993;  77 169-172
• 7 Suzuki Y, Hatanaka Y, Taira M, Shimada F, Hashimoto N, Takayanagi M, Taylor SI, Makino H, Yoshida S. Insulin resistance associated with decreased levels of insulin-receptor messenger ribonucleic acid: evidence of a de novo mutation in the maternal allele.  J Clin Endocrinol Metab. 1995;  80 1214-1220
• 8 Makino H, Taira M, Shimada F, Hashimoto N, Suzuki Y, Nozaki O, Hatanaka Y, Yoshida S. Insulin receptor gene mutation: a molecular genetical and functional analysis.  Cell Signal. 1992;  4 351-363
• 9 Hashimoto N, Zhang WR, Goldstein BJ. Insulin receptor and epidermal growth factor receptor dephosphorylation by three major rat liver protein-tyrosine phosphatases expressed in a recombinant bacterial system.  Biochem J. 1992;  284 ((Pt 2)) 569-576
• 10 Tabiti K, Cui L, Chhatwal VJ, Moochhala S, Ngoi SS, Pallen CJ. Novel alternative splicing predicts a secreted extracellular isoform of the human receptor-like protein tyrosine phosphatase LAR.  Gene. 1996;  175 7-13
• 11 Norris K, Norris F, Kono DH, Vestergaard H, Pedersen O, Theofilopoulos AN, Moller NP. Expression of protein-tyrosine phosphatases in the major insulin target tissues.  FEBS Lett. 1997;  415 243-248
• 12 Seely BL, Staubs PA, Reichart DR, Berhanu P, Milarski KL, Saltiel AR, Kusari J, Olefsky JM. Protein tyrosine phosphatase 1B interacts with the activated insulin receptor.  Diabetes. 1996;  45 1379-1385
• 13 Ahmad F, Goldstein BJ. Functional association between the insulin receptor and the transmembrane protein-tyrosine phosphatase LAR in intact cells.  J Biol Chem. 1997;  272 448-457
• 14 Kenner KA, Anyanwu E, Olefsky JM, Kusari J. Protein-tyrosine phosphatase 1B is a negative regulator of insulin- and insulin-like growth factor-I-stimulated signaling.  J Biol Chem. 1996;  271 19810-19816
• 15 Hashimoto N, Feener EP, Zhang WR, Goldstein BJ. Insulin receptor protein-tyrosine phosphatases. Leukocyte common antigen-related phosphatase rapidly deactivates the insulin receptor kinase by preferential dephosphorylation of the receptor regulatory domain.  J Biol Chem. 1992;  267 13811-13814
• 16 Elchebly M, Payette P, Michaliszyn E, Cromlish W, Collins S, Loy AL, Normandin D, Cheng A, Himms-Hagen J, Chan CC, Ramachandran C, Gresser MJ, Tremblay ML, Kennedy BP. Increased insulin sensitivity and obesity resistance in mice lacking the protein tyrosine phosphatase-1B gene.  Science. 1999;  283 1544-1548
• 17 Zabolotny JM, Kim YB, Peroni OD, Kim JK, Pani MA, Boss O, Klaman LD, Kamatkar S, Shulman GI, Kahn BB, Neel BG. Overexpression of the LAR (leukocyte antigen-related) protein-tyrosine phosphatase in muscle causes insulin resistance.  Proc Natl Acad Sci USA. 2001;  98 5187-5192
• 18 Dadke S, Kusari A, Kusari J. Phosphorylation and activation of protein tyrosine phosphatase (PTP) 1B by insulin receptor.  Mol Cell Biochem. 2001;  221 147-154
• 19 Hauguel-de Mouzon S, Peraldi P, Alengrin F, Obberghen E Van. Alteration of phosphotyrosine phosphatase activity in tissues from diabetic and pregnant rats.  Endocrinology. 1993;  132 67-74
• 20 Ukkola O, Santaniemi M. Protein tyrosine phosphatase 1B: a new target for the treatment of obesity and associated co-morbidities.  J Intern Med. 2002;  251 467-475

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