Precision targeting of protein kinases. An affinity label that inactivates the cGMP- but not the cAMP-dependent protein kinase.
Although the cAMP-dependent (PKA) and cGMP-dependent protein kinases (PKG) usually participate in unrelated biological processes, their enzymological properties are decidedly similar. Based upon the multitude of comparative studies conducted to date, it appears that these two enzymes exhibit very similar peptide substrate specificities. Furthermore, most inhibitors that have been reported for PKG serve in a nearly equal capacity for PKA. Consequently, the task of distinguishing between these enzymes, especially under in vivo conditions, has proved to be daunting. However, we have recently found that PKA will only phosphorylate non-amino acid residues whose alpha-configuration corresponds to that found in L-amino acids, whereas PKG will catalyze the phosphorylation of residues corresponding to both L- and D-amino acids (Wood, J., Mendelow, M., Yan, X., Corbin, J.D., Francis, S.H., and Lawrence, D.S. (1996) J. Biol. Chem. 271, 174-179). Based on these results, we have designed a potent affinity label for PKG (KI = 21.1 +/- 4.7 microM), that has no measurable activity toward PKA. This represents the first example of an peptide-based inactivator that fully distinguishes between these two closely related enzymes. These results suggest that a similar strategy may provide highly specific inactivators for other protein kinases as well.
Yan
X
X
Department of Chemistry, State University of New York, Buffalo 14260, USA.
Corbin
J D
JD
Francis
S H
SH
Lawrence
D S
DS
D000255:Adenosine Triphosphate / Q000378:metabolism; D000345:Affinity Labels; D000595:Amino Acid Sequence; D001665:Binding Sites; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry*; D017869:Cyclic GMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors*; D003545:Cysteine / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000737:chemistry*; D007700:Kinetics; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry; D013379:Substrate Specificity
D000345:Affinity Labels; D004791:Enzyme Inhibitors; D009842:Oligopeptides; D000255:Adenosine Triphosphate; D017868:Cyclic AMP-Dependent Protein Kinases; D017869:Cyclic GMP-Dependent Protein Kinases; D003545:Cysteine