Boyd Haley | University of Kentucky College of Arts & Sciences

Boyd Haley

Professor Emeritus

Contact Information

behaley@ctiscience.com

Affiliations

Chemistry

Research

The basic research interest of our laboratory centers on biochemical and biomedical problems involving control at the molecular level. Specifically, we are most interested in biological systems regulated by protein-nucleotide interactions where the bioenergetics involved are expressed through site-specific nucleotide binding of high affinity or through protein substrate phosphorylation. Our approach to this general phenomenon is to synthesize novel nucleotide analogs that are photoactive or fluorescent, or both. The analogs are then used to study various aspects of protein-nucleotide interactions which regulate enzyme activity. Analogs used may be modifications of the commonly known nucleotides such as ATP, cAMP, GTP, dUTP, NAD+, UDP-Glc, etc. or probes of the more unusual nucleotides such as the proposed alamones guanosine-3 -phosphate-5 -phosphate (magic spot compounds) and diadenosine-P1, P4-tetraphosphate.

These nucleotide analogs are used to study a wide variety of enzymes that either use or are regulated by nucleotides including cyclases, kinases, and polymerases. Certain of these probes have proven useful in the study of the differences between normal and diseased tissues as observed through changes in nucleotide binding proteins. Alzheimer's disease and colon carcinoma are two of the diseases we are currently investigating. Additionally, we have found that several of the large polypeptide hormones (or biological response modifiers) have nucleotide binding sites. These include IL-1, IL-2, Interferon- alpha, TNF (tumor necrosis factor), glucagon and GH (growth hormone) from several sources. We therefore feel that the transmembrane signaling effected by these proteins may, at least partially, be expressed partially by an enzyme activity associated with the "hormone" itself. This is supported by the observation that IL-1 and IL-2 also autophosphorylate using ATP.

Graduate Training

Bioorganic Chemistry

Selected Publications

Selected Publications:

R. R. Drake, R. K. Evans, M. J. Wolf and B. E. Haley, "Synthesis and Properties of 5-Azido-UDP-Glucose: Development of Photoaffinity Probes for Nucleotide Diphosphate Sugar Binding Sites," J. Biol. Chem., 264, 11928-11933 (1989).
A. J. Chavan, H. Kim, B. E. Haley and D. S. Watt, "A Photoactive Phosphonamide Derivative of GTP for the Identification of the GTP Binding Domain of beta-Tubulin", Bioconjugate Chemistry, 1, No. 5, 337-344 (1990).
S. Campbell, H. Kim, M. Doukas and B. Haley, "Photoaffinity Labeling of ATP and NAD+ Binding Sites on Recombinant Human Interleukin-2," Proc. Natl. Acad. Sci., 87, 1243-1246 (1990).
H. Kim and B. Haley, "Identification of Peptides in the Adenine Ring Binding Domain of Glutamate and Lactate Dehydrogenase Using 2-Azido-NAD+," Bioconjugate Chemistry, 2, 142-147 (1991).
B. Haley, "Nucleotide Photoaffinity Labeling and Identification of Protein Kinase Subunits," Meth. Enzymol., 200, 477-487 (1991).
M. Salvucci, A. Chavan and B. Haley, "Identification of Peptides for the Adenine Binding domains of ATP and AMP in Adenylate Kinase: Isolation of Photoaffinity Labeled Peptides by Metal Chelate Chromatography," Biochemistry, 31, 4479-4487 (1992).
M. Shoemaker, P. C. Lin and B. Haley, "Identification of the Guanine Binding Domain Peptide of the GTP Binding Site of Glucagon," Protein Science, 1, 884-891 (1992).
A. Chavan, Y. Nemoto, S. Narumiya, S. Kozaki and B. Haley, "NAD+ Binding Site of Clostridium Botulinum C3 ADP-ribosyltransferase: Identification of Peptide in the Adenine Ring Binding Domain Using 2-Azido NAD+," J. Biol. Chem., 267, 14866-14870 (1992).
M. Doukas, A. Chavan, C. Gass, T. Boone and B. Haley, "Identification and Characterization of a Nucleotide Binding Site on Recombinant Murine Granulocyte/Macrophase-Colony Stimulating Factor. Bioconjugate Chemistry, in press (1992).
D. J. Gunnersen and B. E. Haley, "Detection of glutamine Synthetase in the Cerbrospinal Fluid of Alzheimer's Diseased Patients: A Potential Diagnostic Biochemical Marker," Proc. Natl. Acad. Sci. USA, accepted (1992).
M. Shoemaker and B. E. Haley, "Identification of the Guanine Binding Domain Within Glutamate Dehydrogenate," submitted (1992).
D. M. Hiestand, B. E. Haley and M. S. Kindy, "Role of Calcium in Inactivation of Calcium/-Calmodulin Dependent Protein Kinase II After Cerebral Ischemia," Journal of the Neurological Sciences, 113, 31-37 (1992).
M. E. Salvucci, A. J. Chavan and B. E. Haley, "Identification of Peptides from the Adenine Binding Domains of ATP and AMP in Adenylate Kinase: Isolation of Photoaffinity-Labeled Peptides by Metal Chelate Chromatography," Biochemistry, 31, 4479-4487 (1992).
M. Shoemaker, P. C. Lin and B. Haley, "Identification of the Guanine Binding Domain Peptide of the GTP-Binding Site of Glucagon," Protein Science, 1, 884-891 (1992).
M. A. Doukas, A. J. Chavan, C. Gass, T. Boone and B. E. Haley, "Identification and Characterization of a Nucleotide Binding Site on Recombinant Murine Granulocyte/-Macrophage-Colony Stimulating Factor," Bioconjugate Chemistry, 3, 484-492 (1992).
A. W. Segal, I. West, F. Wientjes, J. H. A. Nugent, A. J. Chavan, B. Haley, R. C. Garcia, H. Rosen and G. Scrace, "Cytochrome b 245 is a Flavocytochrome Containing FAD and the NADPH-Binding Site of the Microbicidal Oxidase of Phagocytes," Biochem. J., 284, 781-788 (1992).
D. C. Hammond, B. E. Haley and J. A. Lesnaw, "Identification and Characterization of Serine/Threonine Protein Kinase Activity Intrinsic to the L Protein of Vesicular Stomatitis Virus New Jersey," Journal of General Virology, 73, 67-75 (1992).
A. J. Chavan, Y. Nemoto, S. Narumiya, S. Kozaki and B. E. Haley, "NAD+ Binding Site of Clostridium botulinum C3 ADP-ribosyltransferase: Identification of Peptide in the Adenine Ring Binding Domain Using 2-Azido NAD+," J. Biol. Chem., 267, 14866-14870 (1992).
D. Gunnersen and B. Haley, "Detection of Glutamine Synthetase in the Cerebrospinal Fluid of Alzheimer's Diseased Patients: A Potential Diagnostic Biochemical Marker," Proc. Natl. Acad. Sci. USA, 89, 11949-11953 (1992).
M. Shoemaker and B. Haley, "Identification of a Guanine Binding Domain Peptide of the GTP Binding Site of Glutamate Dehydrogenase: Isolation with Metal-Chelate Affinity Chromatography," Biochemistry, 32, 1883-1890 (1993).
S. B. Churn, B. Sankaran, B. E. Haley and R. J. Delorenzo, "Ischemic Brain Injury Selectively Alters ATP Binding of Calcium and Calmodulin-Dependent Protein Kinase-II," Biochem. Biophys. Res. Comm., 193(3), 934-940 (1993).
M. Salvucci, K. Rajagopalan, G. Sievert, B. Haley and D. Watt, "Photoaffinity Labeling of Rubisco Activase with ATP- -benzophenone: Identification of the ATP -Phosphate Binding Domain," J. Biol. Chem., 268, 14239-14244 (1993).
K. Rajagopalan, A. Chavan, B. Haley and D. Watt, "Bidentate Cross-Linking Reagents: Non-Hydrolyzable Nucleotide Photoaffinity Probes with Two Photoactive Groups," J. Biol. Chem., 268, 14245-14253 (1993).
C. Trad, A. Chavan, J. Clemens and B. Haley, "Identification and Characterization of an NADH Binding Site of Prolactin with 2-Azido-NAD+," Arch. Biochem. Biophys., 304, 58-64 (1993).
A. Chavan, C. Ensor, P. Wu, B. Haley and H. Tai, "Photoaffinity Labeling of Human Placental NAD+-Linked 15-Hydroxyprostaglandin Dehydrogenase with [ 32P]-2N3NAD+: Identification of a Peptide in the Adenine Ring Binding Domain," J. Biol. Chem., 268, 16437-16442 (1993).
A. Chavan, S. Richardson, H. Kim, B. Haley and D. Watt, "Forskolin Photoaffinity Probes for the Evaluation of Tubulin Binding Sites," Bioconjugate Chem., 4, 268-274 (1993).
E. F. Duhr, J. C. Pendergrass, J. T. Selvin and B. Haley, "HgEDTA Complex Inhibits GTP Interactions with the E-Site of Brain -Tubulin," Toxicology and Applied Pharmacology, 122, 273-288 (1993).
B. Jayaram and B. Haley, "Identification of Peptides Within the Base Binding Domains of the GTP and ATP Specific Binding Sites of Tubulin," J. Biol. Chem., 269(5), 3233-3242 (1994).
A. Chavan, B. Haley, D. Volkin, K. Marfia, A. Verticelli, M. Bruner, J. Draper, C. Burke and R. Middaugh, "Interaction of Nucleotides with Acidic Fibroblast Growth Factor (FGF-1)," Biochemistry, 33, 7193-7202 (1994).
J. Logan, D. Hiestand, P. Daram, Z. Huang, D. Muccio, J. Hartman, B. Haley, W. Cook and E. Sorscher, "Cystic Fibrosis Transmembrane Conductance Regulator Mutations that Disrupt Nucleotide Binding," J. Clin. Invest., 94, 228-236 (1994).
M. Olcott and B. Haley, "Identification of Two Peptides from the ATP-Binding Domain of Creatine Kinase," Biochemistry, 33, 11935-11941 (1994).
A. Bhattacharyya, A. Chavan, M. Shuffett, B. Haley and D. Collins, "Photoaffinity Labeling of Rat Liver Microsomal 5 -Reductase by 2-Azido-NADP+," Steroids, 59, 634-641 (1994).
M. Salvucci, A. Chavan, R. Klein, K. Rajagopalan and B. Haley, "Photoaffinity Labeling of the ATP Binding Domain of Rubisco Activase and a Separate Domain Involved in the Activation of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase," Biochemistry, 33, 14879-14886 (1994).
J. C. Pendergrass and B. E. Haley, "Mercury-EDTA Complex Specifically Blocks Brain -Tubulin-GTP Interactions: Similarity to Observations in Alzheimer's Disease," in Status Quo and Perspective of Amalgam and Other Dental Materials, (International Symposium Proceedings, L. T. Friberg and G. N. Schrauzer, Eds.), Georg Thieme Verlag, Stuttgart-New York, 1995, pp. 98-105.
M. Doukas, A. Chavan, C. Gass, P. Nickel, T. Boone and B. Haley, "Inhibition of GM-CSF Activity by Suramine and Suramin Analogues is Correlated to Interaction with the GM-CSF Nucleotide Binding Site," Cancer Research, 55, 5161-5163 (1995).
A. K. Bhattacharyya, A. J. Chavan, B. Haley, M. F. Taylor and D. C. Collins, "Identification of the NADP(H) Binding Site of Rat Liver Microsomal 5 -Reductase (Isozyme-1): Purification of a Photolabeled Peptide Corresponding to the Adenine Binding Domain," Biochemistry, 34, 3663-3669 (1995).
A. Chavan, C. Gass, B. Haley, T. Boone and M. A. Doukas, "Identification of N-Terminus Peptide of Human Granulocyte/Macrophage Colony Stimulating Factor as the Site of Nucleotide Interaction," Biochem. Biophys. Res. Commun., 208(1), 390-396 (1995).
M. Shoemaker and B. Haley, "Identification of the Adenine Binding Domain Peptides of the ADP Binding Site of Glutamate Dehydrogenase," Bioconjugate Chemistry, 7, 302-310 (1996).
K. Rajagopalan, G. Pavlinkova, S. Levy, R. Pokkuluri, M. Schiffer, B. Haley and H. Kohler," Novel Unconventional Binding Site in the Variable Region of Immunoglobulins," Proc. Natl. Acad. Sci., 93, 6019-6024 (1996).
J. C. Pendergrass and B. E. Haley, "Inhibition of Brain Tubulin-Guanosine 5 -Triphosphate Interactions by Mercury: Similarity to Observations in Alzheimer's Diseased Brain," in Metal Ions in Biological Systems V34, Mercury and Its Effects on Environment and Biology, H. Sigel and A. Sigel, Eds., Marcel Dekker, Inc., 270 Madison Avenue, New York, NY, Chapter 16, 10016 (1996).
M. McGuire, L. J. Carroll, L. Yankie, S. H. Thrall, D. Dunaway-Mariano, O. Hertzberg, B. Jayaram and B. Haley, "Determination of the Nucleotide Binding Site Within Clostridium symbiosum Pyruvate Phosphate Dikinase by Photoaffinity Labeling, Site-Directed Mutagenesis, and Structural Analysis," Biochemistry, 35, 8544-8552 (1996).
H. Kohler, G. Pavlinkova and B. Haley, "Immunoglobulin Nucleotide Binding Site: A Possible Superantigen Receptor," in Human B Cell Superantigens, M. Zouali, Ed., Chapter 13, 1996, pp. 189-194.
B. Sankaran, A. Chavan and B. E. Haley, "Identification of Adenine Binding Domain Peptides of the NADP+ Active Site Within Porcine Heart NADP+-Dependent Isocitrate Dehydrogenase," Biochemistry, 35, 13501-13510 (1996).
G. Pavlinkova, K. Rajagopalan, S. Muller, A. Chavan, G. Sievert, D. Lou, C. O'Tolle, B. Haley and H. Kohler, "Site-Specific Photobiotinylation of Immunoglobins, Fragments and Light Chain Dimers," J. Immunological Methods, 201, 77-88 (1997).
B. Sankaran, J. Clemens and B. Haley, "A Comparison of Changes in Nucleotide-Protein Interactions in Striatal, Hippocampus and Paramedian Cortex After Cerebral Ischemia and Reperfusion: Correlations to Regional Vulnerability," Molecular Brain Research, 47, 237-250 (1997).
K. Hensley, P. Cole, R. Subramaniam, M. Aksenov, M. Aksenova, P. M. Bummer, B. E. Haley, J. M. Carney and D. A. Butterfield, , "Oxidatively-Induced Structural Alteration of Glutamine Synthetase Assessed by Analysis of Spin Labeled Incorporation Kinetics: Relevance to Alzheimer's Disease," J. Neurochem., 68, 2451-2457 (1997).
J. C. Pendergrass, B. E. Haley, M. J. Vimy, S. A. Winfield and F. L. Lorscheider, "Mercury Vapor Inhalation Inhibits Binding of GTP to Tubulin in Rat Brain: Similarity to a Molecular Lesion in Alzheimer's Disease Brain," Neurotoxicology, 18(2), 315-324 (1997).
M. C. Olcott and B. E. Haley, "Identification of an Adenine-Nucleotide Binding Site on Interferon- 2," Eur. J. Biochem., 247(3), 762-769 (1997).
S. David, M. Shoemaker and B. Haley, "Abnormal Properties of Creatine Kinase in Alzhiemer's Disease Brain: Correlation of Reduced Enzyme Activity and Active Site Photolabeling with Aberrant Cytosol-Membrane Partitioning," Molecular Brain Research, 54, 276-287 (1998).
Y. Chen, R. C. Haddon, S. Fang, A. M. Rao, P. C. Eklund, W. H. Lee, E. C. Dickey, E. A. Grulke, J. C. Pendergrass, A. Chavan, B. E. Haley and R. E. Smalley, "Chemical Attachment of Organic Functional Groups to Single-Walled Carbon Nanotube Material," J. Mater. Res., 13, 2423-2431 (1998).
S. S. M. David, and B. E. Haley, "ATP Nucleotidylation of Creatine Kinase", Biochemistry 38, 8492-8500 (1999).
K. Rajagopalan, D. S. Watt, and B. E. Haley, "Orientation of GTP and ADP within their respective binding sites in glutamate dehydrogenase", Eur. J. Biochem. 265, 564-571 (1999).