Substrate specificity and kinetic mechanism of human placental insulin receptor/kinase

Walker, Duncan; Kuppuswamy, Dhandapani; A, Visvanathan; Pike, Linda J.

doi:10.1021/bi00379a033

Cited by 37 publications

(26 citation statements)

References 26 publications

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“…These results are strikingly exemplified by the mammalian Erk-1 and Erk-2 gene products that autophosphorylate on both tyrosine and threonine residues even when expressed in bacteria (39,40 poly(Glu80Tyr20) or insulin receptor-1155-1165) peptide, which is derived from the major insulin receptor tyrosine phosphorylation site (not illustrated). These observations are consistent with the data of Walker et al (41), who examined the hydroxylamino acid selectivity of the insulin receptor tyrosine kinase using the insulin receptor synthetic peptide substrate Arg-Arg-Leu-Ile-Glu-Asp-Ala-Glu-Tyr-Ala-AlaArg-Gly and serine/threonine analogues thereof. In a conventional kinase assay, no phosphorylation of the analogues was detectable.…”

Section: Discussionsupporting

confidence: 92%

Catalysis of serine and tyrosine autophosphorylation by the human insulin receptor.

Baltensperger

Lewis

Woon

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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The protein kinase activity of human insulin receptors purified from Sf9 insect cells after infection with a recombinant baculovirus was evaluated. The following exper-imental observations led to the unexpected conclusion that this receptor protein catalyzes both serine and tyrosine autophos- The insulin receptor is one of a number of growth factor receptors with intrinsic tyrosine kinase activity that can be activated upon binding of appropriate peptide ligands (1). Binding of insulin to its receptor also causes rapid phosphorylation of the tyrosine residues of the insulin receptor (3 subunit itself (2-4), which in turn further activates the tyrosine kinase activity (5, 6). In intact cells, insulin causes receptor phosphorylation on tyrosine as well as serine and threonine residues (2). Protein kinase C or protein kinases activated by this enzyme can phosphorylate the serine and threonine residues of the insulin receptor in intact cells (7,8). Stimulation of serine phosphorylation of the insulin receptor by phorbol esters appears to correlate with inhibition of the insulin-stimulated tyrosine kinase activity ofthe receptor (8).Thus, phosphorylation ofthe serine and threonine residues of the insulin receptor may regulate insulin signaling by modulating receptor tyrosine kinase activity. Several laboratory groups have reported the detection of an insulin-stimulated and receptor-associated serine/ threonine kinase activity using the receptor itself as substrate, in partially purified (9-14) or affinity-purified (15) insulin receptor preparations.

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Section: Discussionsupporting

confidence: 92%

Catalysis of serine and tyrosine autophosphorylation by the human insulin receptor.

Baltensperger

Lewis

Woon

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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“…A suitable peptide substrate product inhibitor is not available that can be used to distinguish between the obligatory initial binding of ATP and random addition of substrates. The catalytic mechanism of the full-length ligand-stimulated activated PDGF ␤-receptor kinase is also found to be a sequential rather than ping-pong mechanism (37), and the epidermal growth factor receptor (38) and insulin receptor kinase (39) reactions are reported to occur by a sequential Bi Bi rapid equilibrium random mechanism. In addition, sequential mechanisms are reported for other tyrosine kinases, such as csk (40) and pp60 src (41), and for the serine/threonine kinases, cAMP-dependent protein kinase (42), and p38 kinase (43), the latter utilizing an ordered sequential mechanism in which the peptide substrate binds first (43).…”

Section: Identification Of the Sequence Of Enzymatically Active Kdrmentioning

confidence: 97%

Vascular Endothelial Growth Factor Receptor KDR Tyrosine Kinase Activity Is Increased by Autophosphorylation of Two Activation Loop Tyrosine Residues

Kendall

Rutledge

Mao

et al. 1999

Journal of Biological Chemistry

117

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Vascular endothelial growth factor is an important physiological regulator of angiogenesis. The function of this endothelial cell selective growth factor is mediated by two homologous tyrosine kinase receptors, fms-like tyrosine kinase 1 (Flt-1) and kinase domain receptor (KDR). Although the functional consequence of vascular endothelial growth factor binding to the Flt-1 receptor is not fully understood, it is well established that mitogenic signaling is mediated by KDR. Upon sequencing several independent cDNA clones spanning the cytoplasmic region of human KDR, we identified and confirmed the identity of a functionally required valine at position 848 in the ATP binding site, rather than the previously reported glutamic acid residue, which corresponds to an inactive tyrosine kinase. The cytoplasmic domain of recombinant native KDR, expressed as a glutathione S-transferase fusion protein, can undergo autophosphorylation in the presence of ATP. In addition, the kinase activity can be substantially increased by autophosphorylation at physiologic ATP concentrations. Mutation analysis indicates that both tyrosine residues 1054 and 1059 are required for activation, which is a consequence of an increased affinity for both ATP and the peptide substrate and has no effect on k cat , the intrinsic catalytic activity of the enzyme. KDR kinase catalyzes phosphotransfer by formation of a ternary complex with ATP and the peptide substrate. We demonstrate that tyrosine kinase antagonists can preferentially inhibit either the unactivated or activated form of the enzyme.

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“…Phosphatidylinositol kinase activity has been observed in immunoprecipitates of the insulin receptor kinase (68,69) and in association with other tyrosine kinase (70)(71)(72). Insulin action on cells also appears to activate one or more phospholipases, including a specific phospholipase C that may generate an inositol glycan compound capable of mimicking several of insulin's actions on cells (73)(74)(75).…”

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confidence: 99%

The insulin receptor and the molecular mechanism of insulin action.

Kahn¹,

White²

1988

J. Clin. Invest.

406

193

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In humans and most other vertebrates, the primary hormone involved in control of blood glucose is insulin. Insulin acts on cells to stimulate glucose, protein, and lipid metabolism, as well as RNA and DNA synthesis, by modifying the activity of a variety of enzymes and transport processes. The importance ofunderstanding insulin action is not limited to simply satisfying the intellectual needs of the curious biochemist or cell biologist. Elucidating the molecular pathways ofinsulin action forms an important cornerstone upon which to unravel the pathogenesis of non-insulin-dependent (type II) diabetes mellitus and a major component of other insulin-resistant states including obesity, uremia, glucocorticoid, and growth hormone excess, as well as a variety of rarer genetic disorders such as leprechaunism, the type A syndrome of insulin resistance, and lipoatrophic diabetes.The actions of insulin at the cellular level are initiated by insulin binding to its plasma membrane receptor (1-4). This receptor is present on virtually all mammalian tissues, although the concentration varies from as few as 40 receptors on circulating erythrocytes to more than 200,000 receptors on adipocytes and hepatocytes. Like the surface membrane receptors for other hormones and growth factors, the insulin receptor serves at least two functions. The first is to recognize the hormone among all other substances in the blood. This is accomplished by binding the hormone with high affinity and a high degree of specificity. The second function is to produce a transmembrane signal that alters intracellular metabolism and mediates the action ofthe hormone. Recognition ofthe insulin molecule by its receptor is a complex molecular event and is closely linked to signal transmission. The binding domain of the insulin molecule is composed of distant portions of the A and B chains, which come together on one surface as a result of three-dimensional folding to form the receptor binding region (5). Among over 200 analogues of insulin studied, there is an almost perfect correlation between receptor-binding affinity and biological effect (1,5,6). No competitive antagonists of insulin action at the receptor level have, as of yet, been uncovered, suggesting that the structural requirements for binding include all the features necessary for biological action. Interestingly, the binding properties of the insulin receptor are better conserved in evolution than are the properties of the insulin molecule itself (7).Address reprint requests to Dr. C. Ronald Kahn, Joslin Diabetes Center, One Joslin Place, Boston, MA 02215.Receivedfor publication 29 June 1988.The insulin receptor is a heterotetrameric glycoprotein consisting of two a-subunits of Mr = 135,000 and two (3-subunits of Mr = 95,000 linked by disulfide bonds to give a ,3-a-a-13 structure (8, 9) (Fig. 1). The a-subunit appears to be entirely extracellular and contains the insulin binding site. This binding site can be affinity labeled using labeled insulin and bifunctional cross-linking agents (8, 9) or photo...

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Substrate specificity and kinetic mechanism of human placental insulin receptor/kinase

Cited by 37 publications

References 26 publications

Catalysis of serine and tyrosine autophosphorylation by the human insulin receptor.

Catalysis of serine and tyrosine autophosphorylation by the human insulin receptor.

Vascular Endothelial Growth Factor Receptor KDR Tyrosine Kinase Activity Is Increased by Autophosphorylation of Two Activation Loop Tyrosine Residues

The insulin receptor and the molecular mechanism of insulin action.

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