Precision Substrate Targeting of Protein Kinases

The cAMP-dependent (PKA) and cGMP-dependent protein kinases (PKG) share a strong primary sequence homology within their respective active site regions. Not surprisingly, these enzymes also exhibit overlapping substrate specificities, a feature that often interferes with efforts to elucidate their distinct biological roles. In this report, we demonstrate that PKA and PKG exhibit dramatically different behavior with respect to the phosphorylation of α-substituted alcohols. Although PKA will phosphorylate only residues that contain an α-center configuration analogous to that found in L-serine, PKG utilizes residues that correspond to both L- and D-serine as substrates. The PKG/PKA selectivity of these substrates is the highest ever reported.

[1]  D. Mochly‐Rosen.  Localization of protein kinases by anchoring proteins: a theme in signal transduction. , 1995, Science.

[2]  D. Lawrence,et al.  The Extraordinary Active Site Substrate Specificity of pp60c−src , 1995, The Journal of Biological Chemistry.

[3]  T. Hunter,et al.  Protein kinases and phosphatases: The Yin and Yang of protein phosphorylation and signaling , 1995, Cell.

[4]  S. M. Van Patten,et al.  Multiple pathway signal tran sduction by the cAMP‐dependent protein kinase , 1994, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[5]  M. Krieger,et al.  cAMP- and cGMP-dependent protein kinase phosphorylation sites of the focal adhesion vasodilator-stimulated phosphoprotein (VASP) in vitro and in intact human platelets. , 1994, The Journal of biological chemistry.

[6]  D. Lawrence,et al.  Phenol kinase activity of the serine/threonine-specific cAMP-dependent protein kinase: steric and electronic effects. , 1994, Biochemistry.

[7]  A. Newton,et al.  Intrapeptide regulation of protein kinase C. , 1994, The Journal of biological chemistry.

[8]  D. Lawrence,et al.  The active site substrate specificity of protein kinase C. , 1994, The Journal of biological chemistry.

[9]  D. Lawrence,et al.  Molecular Basis for the Substrate Specificity of a Serine/Threonine-Specific Protein Kinase , 1993 .

[10]  D. Lawrence,et al.  Stereochemistry specifies the regiochemistry of phosphorylation in two cAMP-dependent protein kinase substrates. , 1993, The Journal of biological chemistry.

[11]  J. Järv,et al.  Substrate specificity of protein kinase C studied with peptides containing D-amino acid residues. , 1993, Journal of biochemistry.

[12]  D. Lawrence,et al.  The cAMP-Dependent Protein Kinase Discriminates between Prochiral Hydroxyl Groups , 1993 .

[13]  D. Lawrence,et al.  The active site substrate specificity of the cAMP-dependent protein kinase. , 1993, The Journal of biological chemistry.

[14]  Susan S. Taylor,et al.  2.2 A refined crystal structure of the catalytic subunit of cAMP-dependent protein kinase complexed with MnATP and a peptide inhibitor. , 1993, Acta crystallographica. Section D, Biological crystallography.

[15]  Nguyen-Huu Xuong,et al.  Crystal structure of the catalytic subunit of cAMP-dependent protein kinase complexed with magnesium-ATP and peptide inhibitor , 1993 .

[16]  R. Huber,et al.  Phosphotransferase and substrate binding mechanism of the cAMP‐dependent protein kinase catalytic subunit from porcine heart as deduced from the 2.0 A structure of the complex with Mn2+ adenylyl imidodiphosphate and inhibitor peptide PKI(5‐24). , 1993, The EMBO journal.

[17]  J. Corbin,et al.  A phenylalanine in peptide substrates provides for selectivity between cGMP- and cAMP-dependent protein kinases. , 1992, The Journal of biological chemistry.

[18]  D. Walsh,et al.  Structural basis for the low affinities of yeast cAMP-dependent and mammalian cGMP-dependent protein kinases for protein kinase inhibitor peptides. , 1992, Biochemistry.

[19]  J. Zheng,et al.  Crystal structure of the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase. , 1991, Science.

[20]  J. Zheng,et al.  Structure of a peptide inhibitor bound to the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase. , 1991, Science.

[21]  J. Corbin,et al.  Substrate- and kinase-directed regulation of phosphorylation of a cGMP-binding phosphodiesterase by cGMP. , 1990, The Journal of biological chemistry.

[22]  B. Kemp,et al.  Protein kinase recognition sequence motifs. , 1990, Trends in biochemical sciences.

[23]  D. Lawrence,et al.  Cryopreservation of the cyclic 3',5'-adenosine monophosphate-dependent protein kinase from bovine cardiac muscle. , 1989, Journal of biochemical and biophysical methods.

[24]  E. Kaiser.  Synthetic Approaches to Biologically Active Peptides and Proteins Including Enzymes , 1989 .

[25]  T. Hunter.  A thousand and one protein kinases , 1987, Cell.

[26]  P. Greengard,et al.  Distinguishing among protein kinases by substrate specificities. , 1987, Biochemistry.

[27]  M. El-Maghrabi,et al.  Synthetic peptides corresponding to the site phosphorylated in 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase as substrates of cyclic nucleotide-dependent protein kinases. , 1986, The Journal of biological chemistry.

[28]  E. Kaiser,et al.  The use of polymer-bound oximes for the synthesis of large peptides usable in segment condensation: synthesis of a 44 amino acid amphiphilic peptide model of apolipoprotein A-1 , 1985 .

[29]  H. Cheng,et al.  Circular dichroic evidence for an ordered sequence of ligand/binding site interactions in the catalytic reaction of the cAMP-dependent protein kinase. , 1985, Biochemistry.

[30]  S. Ferrari,et al.  Distinct structural requirements of Ca2+/phospholipid‐dependent protein kinase (protein kinase C) and cAMP‐dependent protein kinase as evidenced by synthetic peptide substrates , 1985, FEBS letters.

[31]  R. D. Wade,et al.  Guanosine cyclic 3',5'-phosphate dependent protein kinase, a chimeric protein homologous with two separate protein families. , 1984, Biochemistry.

[32]  K. Titani,et al.  Amino acid sequence of the catalytic subunit of bovine type II adenosine cyclic 3',5'-phosphate dependent protein kinase. , 1983, Biochemistry.

[33]  E. Kaiser,et al.  Synthesis of protected peptide segments and their assembly on a polymer-bound oxime: application to the synthesis of a peptide model for plasma apolipoprotein A-I , 1983 .

[34]  W. DeGrado,et al.  Solid-phase synthesis of protected peptides on a polymer-bound oxime: preparation of segments comprising the sequence of a cytotoxic 26-peptide analog , 1982 .

[35]  E. Krebs,et al.  Phosphorylation by guanosine 3':5'-monophosphate-dependent protein kinase of synthetic peptide analogs of a site phosphorylated in histone H2B. , 1982, The Journal of biological chemistry.

[36]  W. DeGrado,et al.  Polymer-bound oxime esters as supports for solid-phase peptide synthesis. The preparation of protected peptide fragments , 1980 .

[37]  E. Krebs,et al.  Comparison of the substrate specificity of adenosine 3':5'-monophosphate- and guanosine 3':5'-monophosphate-dependent protein kinases. Kinetic studies using synthetic peptides corresponding to phosphorylation sites in histone H2B. , 1979, The Journal of biological chemistry.

[38]  T. Lincoln,et al.  Adenosine 3':5'-cyclic monophosphate- and guanosine 3':5'-cyclic monophosphate-dependent protein kinases: possible homologous proteins. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[39]  Y. Nishizuka,et al.  Studies on the sites in histones phosphorylated by adenosine 3':5'-monophosphate-dependent and guanosine 3':5'-monophosphate-dependent protein kinases. , 1976, The Journal of biological chemistry.

[40]  J. Corbin,et al.  Structure and function of cyclic nucleotide-dependent protein kinases. , 1994, Annual review of physiology.

[41]  J. Corbin,et al.  Progress in understanding the mechanism and function of cyclic GMP-dependent protein kinase. , 1994, Advances in pharmacology.

[42]  J. Corbin,et al.  [26] Purification of type Iα and Type Iβ isozymes and proteolyzed type Iβ monomeric enzyme of cGMP-dependent protein kinase from bovine aorta , 1991 .