Donovan C. Haines
Department of Chemistry
Office: CFS 317
Phone: (936) 294-1530
My research involves the structure and mechanism of action of monooxygenase systems, especially those involved in metabolism of natural products, neurotransmitters, drugs, and hormones. This fundamental research has direct medical relevance as aberrant behavior of oxygen utilizing enzymes has been implicated to varying degrees in many neurodegenerative diseases including Amyotrophic Lateral Sclerosis (Lou Gehrig's disease), Parkinson's disease, and Alzheimer's disease. We specificially focus on lipid metabolizing enzymes, especially the fatty acid hydroxylases. These enzymes have been linked to immune function, regulation of blood pressure and other aspects of the vasculature, and play a role in skin formation (CYP4F22 defects cause the rare disease lamellar ichthyosis type 3). To study these enzymes my laboratory employs organic synthesis to produce enzyme inhibitors, biochemistry to measure the effects of the inhibitors and probe enzyme mechanism, molecular biology to probe the effects of changes in protein sequence on enzyme properties, and biophysics via spectroscopy and crystallography to study the structure and physical properties of the enzymes. In addition to using small molecules to study enzymes, we also use enzymes to synthesize and study interesting small molecules on a research (200 mg) scale.
Our major project involves identifying the enzymes involved in acyl homoserine lactone (AHL) metabolism. AHLs are signal molecules used by bacteria to sense their population density. These signals are important in infection, especially by organisms like Pseudomonas aeruginosa which is a major cause of death in cystic fibrosis patients and cause of limb amputation in diabetics and burn victims. We study how the body repsonds to these signals and modifies them to prevent infections from establishing biofilms and/or expressing virulence factors (things that help the bacteria attack our bodies). We have collaborated with John Teiber at the University of Texas Southwestern Medical Center at Dallas in his study of the role of paraoxonase in this quorum quenching infection defense, and study in our lab the role of P450 cytochromes and related enzymes. Other projects in the lab include develepment of molecular tools to study the role of CYP4F22 in lamellar ichthyosis, aspects of the function of CYP46A1 which oxidizes cholesterol and other steroids in the brain, studies of model P450 enzyme structure and function, and creation of biosensors using Synthetic Biology. In colloboration with Biologists and Forensic Scientists, we also work on the use of proteins, lipids, and volatiles to determine time-of-death and analyze decomposition.
Davydov DR, Ponomarev GV, Bobrovnikova-Marjon E, Haines DC, Peterson JA,"Aluminum-substituted heme domain of P450BM-3 (BMP): Introducing a heme-derived fluorescent probe for studies of substrate binding and protein-protein interactions in cytochromes P450.", Biotechnology and Applied Biochemistry, 60, 1, pp. 41-51, 2013.
Marziaz ML, Frazier K, Guidry PB, Ruiz RA, Petrikovics I, Haines DC, "Comparison of brain mitochondrial cytochrome c oxidase activity with cyanide LD(50) yields insight into the efficacy of prophylactics.", Journal of Applied Toxicology, 33, 1, pp. 50-5, 2013.
De Matteis F, Ballou DP, Coon MJ, Estabrook RW, Haines DC,"Peroxidase-like activity of uncoupled cytochrome P450: studies with bilirubin and toxicological implications of uncoupling.", Biochemical Pharmacology, 84, 3, pp. 374-82, 2012.
Haines DC, Hegde A, Chen B, Zhao W, Bondlela M, Humphreys JM, Mullin DA, Tomchick DR, Machius M, Peterson JA, "A single active-site mutation of P450BM-3 dramatically enhances substrate binding and rate of product formation.", Biochemistry, 50, 39, pp. 8333-41, 2011.
Horke S, Witte I, Altenhofer S, Wilgenbus P, Goldeck M, Forstermann U, Xiao J, Kramer GL, Haines DC, Chowdhary PK, Haley RW, and Teiber JF, "Dominant Paraoxonase 2 is downregulated by the Pseudomonas aeruginosa quorum sensing signal N-(3-oxododecanoyl)-L-homoserine lactone and attenuates oxidative stress induced by pyocyanin.", Biochemical Journal, , 426, 1, pp. 73-83, 2010.
Chowdhary, P.K, Stewart, L., Lopez, C., and Haines , D.C. , "A Single Mutation in P450BM-3 Enhances Acyl Homoserine Lactone : Acyl Homoserine Substrate Binding Selectivity Nearly 250-Fold", Journal of Biotechnology, 135, 4, pp. 374-6, 2008.
Teiber, J.F., Horke, S., Haines, D.C., Chowdhary, P.K., Xiao, J., Kramer, G.L., Haley, R.W. and Draganov, D.I., "Dominant Role of Paraoxonases in the Inactivation of the Pseudomonas aeruginosa Quorum Sensing Signal N-(3-Oxododecanoyl)-L-Homoserine Lactone", Infection and Immunity, 76, 6, pp. 2512-9, 2008.
Haines, D.C., Chen, B., Tomchick, D.R., Bondlela, M., Hegde, A., Machius, M., and Peterson, J.A., "Crystal Structure of Inhibitor Bound P450BM-3 Reveals Open Conformation of Substrate Access Channel", Biochemistry, 47, 12, pp. 3662-70, 2008.
Chowdhary, P.K., Keshavan, N., Nguyen, H., Peterson, J.A., Gonzalez, J.E., and Haines, D.C., “Bacillus megaterium CYP102A1 Oxidation of Acyl Homoserine Lactones and Acyl Homoserines”, Biochemistry, 46, 50, pp. 14429-37, 2007.
Hegde, A., Haines, D.C., Bondlela, M., Chen, B., Schaffer, N., Tomchick, D.R., Machius, M., Nguyen, H., Chowdhary, P.K., Stewart, L., Lopez, C., and Peterson, J.A., "Interactions of Substrates at the Surface of P450s Can Greatly Enhance Substrate Potency", Biochemistry, 46, 49, pp. 14010-7, 2007.
Chowdhary, P.K, Alemseghed, M., and Haines, D.C., “Cloning, Expression, and Characterization of a New Self-Sufficient P450: CYP102A5 from Bacillus cereus”, Archives of Biochemistry and Biophysics, 468, 1, pp. 32-43, 2007.
Kitazume, T., Haines, D.C., Estabrook, R.W., Chen, B., and Peterson J.A., "Obligatory Intermolecular Electron Transfer from FAD to FMN in Dimeric P450BM-3", Biochemistry, 46, 42, pp. 11892-901, 2007.
Haines, D.C., " A Role for the Strained Phenylalanine Ring Rotation Induced by Substrate Binding to Cytochrome CYP102A1", Protein and Peptide Letters, 13, 10, pp. 977-80, 2006.
Keshavan, N.D., Chowdhary, P.K., Haines, D.C., and Gonzalez, J.E., "L-Canavanine Made By Alfalfa Interferes With Quorum Sensing in Sinorhizobium meliloti", Journal of Bacteriology, 187, 24, pp. 8427-36, 2005.
Kou, W., Kolla, H.S., Ortiz-Acevedo, A., Haines, D.C., Junker, M., and Dieckmann, G.R. , "Modulation of Zinc- and Cobalt-Binding Affinites Through Changes in the Stability of the Zinc Ribbon Protein L36" , Journal of Biological Inorganic Chemistry, 10, 2, pp. 167-80, 2005.
Wanduragala, S., Wimalasena, D.S., Haines, D.C., Kahol, P.K., and Wimalasena, K., , pH-Induced Alteration and Oxidative Destruction of Heme in Purified Chromaffin Granule Cytochrome b561: Implications for the Oxidative Stress in Catecholaminergic Neurons, Biochemistry, 42, n. 12, pp. 3617-26, 2003.
Wimalasena, D.S., Jayatillake, S.P., Haines, D.C., Wimalasena, K., Plausible Molecular Mechanism for Fumarate Activation and Electron Transfer of the Dopamine Beta-Monooxygenase Reaction, Biochemical Journal, v. 367, pt. 1, pp. 77-85, 2002.
Haines, D.C., Tomchick, D.R., Machius, M., and Peterson, J.A., The Pivotal Role of Water in the Mechanism of P450BM-3, Biochemistry, 40, n. 45, pp. 13456-65, 2001.
Falck, J.R., Reddy, Y.K, Haines, D.C., Reddy, K.M., Krishna, U.M., Graham, S., Murry, B., and Peterson, J.A., Practical, Enantiospecific Syntheses of 14,15-EET and Leukotoixn B (Vernolic Acid), Tetrahedron Letters, 42, 4131-3, 2001.
Haines, D.C., Sevrioukova, I.F., and Peterson, J.A., The FMN-Binding Domain of P450BM-3: Resolution, Reconstitution, and Flavin Analogue Substitution,Biochemistry, 39, n. 31, pp. 9419-29, 2000.
Wimalasena, K., Wimalasena, D.S., Dharmasena, S., Haines, D.C., and Alliston, K.R., Chiral Multisubstrate Inhibitors of Dopamine b -Monooxygenase: Evidence for Dual Modes of Interaction, Biochemistry, 36, pp. 7144-53. 1997.
Wimalasena, K. and Haines, D.C., A General Progress Curve Method for the Kinetic Analysis of Suicide Enzyme Inhibitors, Analytical Biochemistry, 234, pp. 175-82, 1996.
Wimalasena, K. and Haines, D.C., Nucleophilic Substitution Reactions of Phenacyl Bromide Oxime: Effect of the Solvent and the Basicity of the Nucleophile, Journal of Organic Chemistry, 59, pp. 6472-4, 1994.
Last modified 5/26/2013 .