Design of a potent peptide inhibitor of the epidermal growth factor receptor tyrosine kinase utilizing sequences based on the natural phosphorylation sites of phospholipase C-gamma 1.
Peptides that possess primary sequences identical to segments surrounding the natural phosphorylation sites of phospholipase C-gamma 1 (i.e., tyrosines 472, 771, 783, and 1284) have been synthesized and evaluated with respect to substrate kinetics for the epidermal growth factor receptor tyrosine kinase. A peptide that was based on tyrosine 472 was the superior substrate in terms of lowest Km value at 37 microM and had the following amino acid sequence: Lys-His-Lys-Lys-Leu-Ala-Glu-Gly-Ser-Ala-Tyr472-Glu-Glu-Val. This peptide sequence was used as a foundation to make amino acid substitutions and/or chemical modifications directed toward the synthesis of a potent peptide inhibitor. As a result, a nine amino acid peptide was synthesized having a K(i) of 10 microM.
Fry
D W
DW
Department of Cancer Research, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, MI 48105.
McMichael
A
A
Singh
J
J
Dobrusin
E M
EM
McNamara
D J
DJ
D000595:Amino Acid Sequence; D015195:Drug Design; D066246:ErbB Receptors / Q000037:antagonists & inhibitors*; D007527:Isoenzymes / Q000494:pharmacology*; D007700:Kinetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000494:pharmacology*; D051966:Phospholipase C gamma; D010766:Phosphorylation; D011485:Protein Binding; D013329:Structure-Activity Relationship; D010738:Type C Phospholipases / Q000494:pharmacology*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000494:pharmacology
D007527:Isoenzymes; D010446:Peptide Fragments; C090828:2,3,5,6-tetrafluorotyrosine; D014443:Tyrosine; D066246:ErbB Receptors; D010738:Type C Phospholipases; D051966:Phospholipase C gamma