Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose

doi:10.1016/0092-8674(86)90786-5

Cell

Volume 45, Issue 5, 6 June 1986, Pages 721-732

https://doi.org/10.1016/0092-8674(86)90786-5 Get rights and content

Abstract

Insulin stimulates the autophosphorylation of tyrosine residues of the β subunit of the insulin receptor (IR); this modified insulin-independent kinase has increased activity toward exogenous substrates in vitro. We show here that replacement of one or both of the twin tyrosines (residues 1162 and 1163) with phenylalanine results in a dramatic reduction in or loss of insulin-activated autophosphorylation and kinase activity in vitro. In vivo, these mutations not only result in a substantial decrease in insulin-stimulated IR autophosphorylation but also in a parallel decrease in the insulin-activated uptake of 2-deoxyglucose. Furthermore, a truncated IR protein (lacking the last 112 amino acids) has an unstable β subunit; this mutant has no kinase activity in vitro or in vivo and does not mediate insulin-stimulated uptake of 2-deoxyglucose. IR autophosphorylation is thus implicated in the regulation of IR activities, with tyrosines 1162 and 1163 as major sites of this regulation.

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A strong signal was observed at the expected molecular weight (~100 kDa) in WT, but not in Insr1345H/H KI mice (Figure 2A). Phosphorylation of Y1179 and Y1180, a pair of residues required for insulin-activated kinase activity and uptake of 2-deoxyglucose [5], was not consistently different in the livers of Insr1345H/H mouse when compared with those of WT animals (Figure 2A). Phosphorylation of AKT, a key downstream of mediator of insulin signaling [21], was similar in WT and Insr1345H/H KI mice (Figure 2A).
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Once insulin binds to the extracellular α-chains of the insulin receptor (InsR), the receptor undergoes autophosphorylation of tyrosine residues at Tyr1158, Tyr1162, and Tyr1163 in the activating loop (A-loop) of the insulin receptor kinase (IRK) domain located in the cytoplasmic portion of the β-chains. The subsequent conformational change of the IRK segment enhances kinase activity (Ellis et al., 1986; Rosen, Herrera, Olowe, Petruzzelli, & Cobb, 1983). In the presence of increased insulin receptor serine phosphorylation results in decreased IRK activity (Dunaif, Xia, Book, Schenker, & Tang, 1995; Takayama, White, & Kahn, 1988) or the lack of insulin-stimulation, the un-phosphorylated Tyr1162 residues of the A-loop may compete with ATP and protein substrates to bind at the active site of the kinase domain to lead to an overall loss of phosphorylation and inhibition of full kinase activation (Hubbard, Wei, Ellis, & Hendrickson, 1994).
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Oxidative stress plays a significant role in the development of insulin resistance; however, the cellular targets of oxidation that cause insulin resistance have yet to be fully elucidated. Methionine sulfoxide reductases reduce oxidized methionine residues, thereby repairing and protecting proteins from oxidation. Recently, several genome-wide analyses have found human obesity to be strongly correlated with polymorphisms near the methionine sulfoxide reductase A (MsrA) locus. In this study, we tested whether modulation of MsrA expression significantly alters the development of obesity and/or insulin resistance in mice. We show that mice lacking MsrA (MsrA^−/−) are prone to the development of high-fat-diet-induced insulin resistance and a reduced physiological insulin response compared to high-fat-fed wild-type mice. We also show that oxidative stress in C2C12 cell cultures reduces both insulin-stimulated phosphorylation and autophosphorylation of the insulin receptor. Tissues from high-fat-fed mice show similar reduction in insulin receptor function and increase in insulin receptor oxidation, which are further exacerbated by the lack of MsrA. Together, these data demonstrate for the first time that MsrA and protein oxidation play a role in the regulation of glucose homeostasis. In addition, these data support a novel hypothesis that obesity-induced insulin resistance is caused in part by reduced function of insulin signaling proteins arising from protein oxidation.

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^†: Present address: 17, rue du Fer a Moulin, 75005 Paris, France.

^§: Present address: Laboratoire de Physiologie de la Lactation, INRA CNRZ, 78350 Jouy en Josas, France.

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Cell

ArticleReplacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose

Abstract

J. Biol. Chem.

FEBS Lett.

J. Biol. Chem.

J. Biol. Chem.

Cell

J. Biol. Chem.

J. Mol. Biol.

J. Biol. Chem.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Cell

J. Biol. Chem.

Cell

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Cellular oncogenes and retroviruses

Ann. Rev. Biochem.

Changes in protein phosphorylation in Rous Sarcoma Virus-transformed chicken embryo cells

Mol. Cell. Biol.

Redesigning trypsin: alteration of substrate specificity

Science

Local mutagenesis of Rous sarcoma virus: the major sites of tyrosine and serine phosphorylation of p60arc are dispensible for transformation

Cell

The nature and regulation of the insulin receptor: structure and function

Ann. Rev. Physiol.

Autophosphorylation sites on the epidermal growth factor receptor

Nature

Expression of a functional human insulin receptor from a cloned cDNA in Chinese hamster ovary cells

Stimulation of tyrosine-specific phosphorylation by platelet-derived growth factor

Nature

Site directed mutagenesis shows that tyrosine 248 of carboxypeptidase A does not play a crucial role in catalysis

Nature

Article
Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose

Local mutagenesis of Rous sarcoma virus: the major sites of tyrosine and serine phosphorylation of p60^arc are dispensible for transformation