J. Biol. Chem., Vol. 265, Issue 27, 16205-16209, Sep, 1990

A rationale for the design of an inhibitor of tyrosyl kinase

CJ Yuan, S Jakes, S Elliott and DJ Graves
Department of Biochemistry and Biophysics, Iowa State University, Ames 50011.

Two gastrin analogs containing a D- and a L-tetrafluorinated tyrosyl residue (Arg-Arg-Leu-Glu-Glu-Glu-Glu-Glu-Ala-(F4)Tyr-Gly) were synthesized and tested as substrates and inhibitors of the insulin receptor kinase. No phosphorylation of these peptides was observed, but both gastrin analogs were effective inhibitors in the microM range. Although the D- and L-tetrafluorotyrosine-gastrin analogs differ in the sequence by only 1 amino acid residue, a different inhibitory pattern was obtained with the insulin receptor. The inhibition of all-L-isomer is competitive with respect to both the protein substrate, reduced, S- carboxymethylated, and maleylated lysozyme (RCMM-lysozyme), and ATP with a Ki value of 4 microM. This result corroborates a previous finding (Walker, D. H., Kuppuswamy, D., Visvanathan, A., and Pike, L. J. (1987) Biochemistry 26, 1428-1433) that the kinetic mechanism for insulin receptor is a random Bi Bi mechanism. Different from the L- isomer, the D-analog is competitive to RCMM-lysozyme and noncompetitive toward ATP and gives an apparent inhibition constant of 20 microM. A free tetrafluorotyrosine also shows a competitive inhibition to protein substrate, RCMM-lysozyme (Ki = 18 mM) whereas free tyrosine shows no effect on the activity of insulin receptor. These results show the importance of the charge state and nucleophilicity of the phenolic component in substrate recognition and catalysis and provide a rationale for the design of inhibitors of tyrosyl phosphorylation.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. Niu and D. S. Lawrence Nonphosphorylatable Tyrosine Surrogates. IMPLICATIONS FOR PROTEIN KINASE INHIBITOR DESIGN J. Biol. Chem., January 17, 1997; 272(3): 1493 - 1499. [Abstract] [Full Text] [PDF]

				P. A. Cole, M. R. Grace, R. S. Phillips, P. Burn, and C. T. Walsh The Role of the Catalytic Base in the Protein Tyrosine Kinase Csk J. Biol. Chem., September 22, 1995; 270(38): 22105 - 22108. [Abstract] [Full Text] [PDF]

				A. J. Ablooglu, J. H. Till, K. Kim, K. Parang, P. A. Cole, S. R. Hubbard, and R. A. Kohanski Probing the Catalytic Mechanism of the Insulin Receptor Kinase with a Tetrafluorotyrosine-containing Peptide Substrate J. Biol. Chem., September 22, 2000; 275(39): 30394 - 30398. [Abstract] [Full Text] [PDF]

				D. A. Enke, P. Kaldis, and M. J. Solomon Kinetic Analysis of the Cyclin-dependent Kinase-activating Kinase (Cak1p) from Budding Yeast J. Biol. Chem., October 20, 2000; 275(43): 33267 - 33271. [Abstract] [Full Text] [PDF]

				A. A. Profit, T. R. Lee, J. Niu, and D. S. Lawrence Molecular Rulers: An Assessment of Distance and Spatial Relationships of Src Tyrosine Kinase SH2 and Active Site Regions J. Biol. Chem., March 16, 2001; 276(12): 9446 - 9451. [Abstract] [Full Text] [PDF]