Lawrence Marnett
University Distinguished Professor of Chemistry & Biochemistry
Mary Geddes Stahlman Professor of Cancer Research
Organic, Bioorganic and Biochemistry
Our research group has interests in the areas of protein structure and function, nucleic acid chemistry, drug design and synthesis, and chemical genomics. Much of our work focuses on the biochemistry and molecular biology of oxidation of natural and synthetic chemicals. Areas of interest to us include: mechanisms of oxidation of arachidonic acid and endocannabinoids by cyclooxygenase and lipoxygenase enzymes, design, synthesis, and biochemical evaluation of lipoxygenase and selective cyclooxygenase-2 (COX-2) inhibitors; chemistry and biology of DNA damage by lipid oxidation products; and endogenous pathways of DNA damage in the genesis of human cancer.
The following studies are underway in our labs:
Structure and Function of Fatty Acid Oxygenases
Our laboratory has a long-standing interest in enzymes of arachidonic acid oxygenation. This includes lipoxygenases, which incorporate one molecule of O2 into the carbon framework and cyclooxygenases, which incorporate two molecules of O2. The products of both pathways of oxygenation are substrates for metabolizing enzymes that generate a panoply of lipid mediators. Leukotrienes and prostaglandins are involved in multiple physiological and pathophysiological events, and inhibition of their action is the molecular basis for the pharmacological activities of several important drugs. Foremost among these are non-steroidal anti-inflammatory drugs (NSAIDs) and cyclooxygenase-2 (COX-2) inhibitors. We have conducted extensive functional studies with lipoxygenases and cyclooxygenases based on available crystal structures and employing exhaustive site-directed mutagenesis.
Design, Synthesis, and Evaluation of Novel COX-2 Inhibitors
Cyclooxygenase-2 (COX-2) is the molecular target of non-steroidal anti-inflammatory drugs (NSAIDs) and selective COX-2 inhibitors. Our laboratory has combined structural analysis with functional studies to define the molecular determinants of the interaction of ligands (substrates and inhibitors) with COX-2. For example, we recently reported the identification of a critical H-bonding interaction that leads to the selectivity of aspirin for acetylation of Ser-530 in COX-2. Many NSAIDs are aralkyl carboxylic acids. Comparative analysis of the effect of site-directed mutation of active site residues on the binding of substrates and inhibitors to COX-1 and COX-2 led us to hypothesize that neutral derivatives of esters and amides would bind selectively to COX-2. We tested this hypothesis by synthesizing a series of neutral derivatives of NSAIDs and demonstrating increases in selectivity for COX-2 of several orders of magnitude. We are exploiting this discovery to prepare novel COX-2 inhibitors as anti-inflammatory drugs and cancer preventive agents.
Chemistry and Biology of Endogenous DNA Damage by Lipid Peroxidation Products
Our laboratory has focused on DNA damage by aldehydes produced endogenously in mammalian cells as a result of lipid peroxidation. Malondialdehyde is the major mutagenic product of lipid peroxidation and is produced ubiquitously in animal and human tissues. It reacts with DNA to form a series of adducts that we and others have identified. The major adduct is a pyrimidopurinone that we have abbreviated M1G. This adduct possesses a blocked Watson-Crick base-pairing region so it is expected to be mutagenic. We have evaluated its mutagenicity by synthesizing viral genomes containing M1G at defined positions. Following transfection into bacterial or mammalian hosts, the replicated genome is interrogated to determine the outcome of replication at the site of the adduct. These experiments indicate that M1G is indeed mutagenic. We have used a variation of this approach to establish that M1G is repaired by nucleotide excision repair. To support and extend these in vivo studies, we conduct experiments utilizing adduct-containing duplex DNA molecules or template-primers as substrates for purified DNA polymerases or repair enzymes. These investigations provide more detailed information about the structural and functional basis for induction of mutation. Our laboratory has had a long-standing collaboration with the Stone laboratory in the Chemistry Department at Vanderbilt, which has provided precise information about the structural perturbations introduced into DNA by adducts such as M1G.
Signal Transduction by Lipid Mediators and Lipid Peroxidation Products
A relatively new area of research in the laboratory is the definition of signal transduction pathways stimulated or interrupted by lipid mediators or lipid peroxidation products. The work on lipid mediators is focused on endocannabinoid oxygenation products of COX-2 and 15-lipoxygenase whereas the work on lipid peroxidation products is focused on malondialdehyde, 4-hydroxynonenal, and structurally related molecules. We are particularly interested in events important in controlling the growth and metastasis of cancer cells such as proliferation, migration, apoptosis and angiogenesis.
Each of the projects utilizes a range of the outstanding core facilities available at Vanderbilt (microarray, proteomics, molecular recognition, and high throughput screening) and involves stimulating collaborations with colleagues at Vanderbilt and elsewhere.
Specializations
VICB
Organic Chemistry
Medicinal Chemistry
Chemical Biology
Bioorganic Chemistry
Biochemistry
Representative Publications
Ra H, González-González E, Uddin MJ, King BL, Lee A, Ali-Khan I, Marnett LJ, Tang JY, Contag CH. Detection of Non-Melanoma Skin Cancer by in vivo Fluorescence Imaging with Fluorocoxib A. Neoplasia. 2015, 17(2):201-7.
Aluise CD, Camarillo JM, Shimozu Y, Galligan JJ, Rose KL, Tallman KA, Marnett LJ. Site-Specific, Intramolecular Cross-Linking of Pin1 Active Site Residues by the Lipid Electrophile 4-Oxo-2-nonenal. Chemical Research in Toxicology. 2015.
Kudalkar SN, Nikas SP, Kingsley PJ, Xu S, Galligan JJ, Rouzer CA, Banerjee S, Ji L, Eno MR, Makriyannis A, Marnett LJ.13-methylarachidonic Acid is a positive allosteric modulator of endocannabinoid oxygenation by cyclooxygenase. The Journal of Biological Chemistry. 2015. 290(12):7897-909.
Xu S, Rouzer CA, Marnett LJ.Oxicams, a class of nonsteroidal anti-inflammatory drugs and beyond. IUBMB Life. 2014, 66(12):803-11.
Shonesy BC, Bluett RJ, Ramikie TS, Báldi R, Hermanson DJ, Kingsley PJ, Marnett LJ, Winder DG, Colbran RJ, Patel S. Genetic disruption of 2-arachidonoylglycerol synthesis reveals a key role for endocannabinoid signaling in anxiety modulation . Cell Reports. 2014, 9(5):1644-53
DeGuire SM, Earl DC, Du Y, Crews BA, Jacobs AT, Ustione A, Daniel C, Chong KM, Marnett LJ, Piston DW, Bachmann BO, Sulikowski GA. Fluorescent probes of the apoptolidins and their utility in cellular localization studies. Angewandte Chemie International Ed Engl. 2015, 54(3):961-4.
Shuck SC, Rose KL, Marnett LJ. Mass spectrometric methods for the analysis of nucleoside-protein cross-links: application to oxopropenyl-deoxyadenosine. Chemical Research in Toxicology. 2014, 27 (1): 136-46.
Xu S, Hermanson DJ, Banerjee S, Ghebreselasie K, Clayton GM, Garavito RM, Marnett LJ. Oxicams Bind in a Novel Mode to the Cyclooxygenase Active Site via a Two-water-mediated H-bonding Network. The Journal of Biological Chemistry. 2014, 0 (0): [Epub ahead of print].
Codreanu SG, Ullery JC, Zhu J, Tallman KA, Beavers WN, Porter NA, Marnett LJ, Zhang B, Liebler DC. Alkylation damage by lipid electrophiles targets functional protein systems. Molecular & Cellular Proteomics.2014, (3): 849-59.
Uddin MJ, Crews BC, Ghebreselasie K, Marnett LJ. Design, synthesis, and structure-activity relationship studies of fluorescent inhibitors of cycloxygenase-2 as targeted optical imaging agents. Bioconjugate Chemistry. 2013, 24 (4): 712-23.
Aldrovandi M, Hammond VJ, Podmore H, Hornshaw M, Clark SR, Marnett LJ, Slatter DA, Murphy RC, Collins PW, O'Donnell VB. Human platelets generate phospholipid-esterified prostaglandins via cyclooxygenase-1 that are inhibited by low dose aspirin supplementation. Nature Neuroscience. 2013, 54 (11): 3085-97.
Hermanson DJ, Hartley ND, Gamble-George J, Brown N, Shonesy BC, Kingsley PJ, Colbran RJ, Reese J, Marnett LJ, Patel S. Substrate-selective COX-2 inhibition decreases anxiety via endocannabinoid activation. Nature Neuroscience. 2013, 16(9): 1291-8.
Liu Q, Ullery J, Zhu J, Liebler DC, Marnett LJ, Zhang B. RNA-seq data analysis at the gene and CDS levels provides a comprehensive view of transcriptome responses induced by 4-hydroxynonenal. Molecular BioSystems2013, 9 (12): 3036-46.
Windsor MA, Valk PL, Xu S, Banerjee S, Marnett LJ. Exploring the molecular determinants of substrate-selective inhibition of cyclooxygenase-2 by lumiracoxib. Bioorganic & Medicinal Chemistry Letters. 2013, 23 (21): 5860-4.
Maddukuri L, Shuck SC, Eoff RL, Zhao L, Rizzo CJ, Guengerich FP, Marnett LJ. Replication, Repair, and Translesion Polymerase Bypass of N(6)-Oxopropenyl-2'-deoxyadenosine. Biochemistry. 2013, 52(48): 8766-76.
Cekanova M, Uddin MJ, Bartges JW, Callens A, Legendre AM, Rathore K, Wright L, Carter A, Marnett LJ. Molecular Imaging of Cyclooxygenase-2 in Canine Transitional Cell Carcinomas In Vitro and In Vivo. Cancer Prevention Research. 2013, 6 (5): 466-476.
Liedtke AJ, Adeniji AO, Chen M, Byrns MC, Jin Y, Christianson DW, Marnett LJ. Penning TM. Development of Potent and Selective Indomethacin Analogues for the Inhibition of AKR1C3 (Type 5 17 beta-Hydroxysteroid Dehydrogenase/Prostaglandin F Synthase) in Castrate-Resistant Prostate Cancer 7.Journal of Medicinal Chemistry. 2013, 56 (6): 2429-2446.
Boiani M, Daniel C, Liu XY, Hogarty MD, Marnett LJ. The Stress Protein BAG3 Stabilizes Mcl-1 Protein and Promotes Survival of Cancer Cells and Resistance to Antagonist ABT-737. Journal of Biological Chemistry. 2013, 288 (10): 6980-6990.
Tallman KA, Armstrong MD, Milne SB, Marnett LJ, Brown HA, Porter NA. Cobalt carbonyl complexes as probes for alkyne-tagged lipids. Journal of Lipid Research.2013, 25 (2): 859-868.
Aluise CD, Rose K, Boiani M, Reyzer ML, Manna JD, Tallman K, Porter NA, Marnett LJ. Peptidyl-prolyl cis/trans-Isomerase A1 (Pin1) Is a Target for Modification by Lipid Electrophiles. Chemical Research in Toxicology. 2013, 26 (2): 270-279.
Yang Y, Wang Z, Varadaraj K, Kumari SS, Mergler S, Okada Y, Saika S, Kingsley PJ, Marnett LJ, Reinach PS. Cannabinoid receptor 1 suppresses transient receptor potential vanilloid 1-induced inflammatory responses to corneal injury. Cellular Signalling. 2013, 25 (2): 501-511.
Sevick-Muraca EM, Akers WJ, Joshi BP, Luker GD, Cutler CS, Marnett LJ, Contag CH, Wang TD, Azhdarinia A. Advancing the translation of optical imaging agents for clinical imaging. Biomedical Optics Express.2013, 4 (1): 160-170.
Scholz M, Blobaum AL, Marnett LJ, Hey-Hawkins E. ortho-Carbaborane derivatives of indomethacin as cyclooxygenase (COX)-2 selective inhibitors. Bioorganic & Medicinal Chemistry. 2012, 20 (15): 4830-4837.
Musee J, Marnett LJ. Prostaglandin H Synthase-2-catalyzed Oxygenation of 2-Arachidonoylglycerol Is More Sensitive to Peroxide Tone than Oxygenation of Arachidonic Acid. Journal of Biological Chemistry. 2012, 287 (44): 37383-37394.
Xu S, Mueser TC, Marnett LJ, Funk MO. Crystal Structure of 12-Lipoxygenase Catalytic-Domain-Inhibitor Complex Identifies a Substrate-Binding Channel for Catalysis. Structure. 2012, 20 (9): 1490-1497.
Marnett LJ. Inflammation and Cancer: Chemical Approaches to Mechanisms, Imaging, and Treatment. Journal of Organic Chemistry. 2012, 77 (12): 5224-5238.
Eisenbrand G, Gates KS, Hecht SS, Janzowski C, Keefer LK, Marnett LJ, Tannenbaum SR. Richard Loeppky (1937-2012) Obituary. Chemical Research in Toxicology. 2012, 25 (6): 1155-1156.
Aldrich MB, Marshall MV, Sevick-Muraca EM, Lanza G, Kotyk J, Culver J, Wang LHV, Uddin J, Crews BC, Marnett LJ, Liao JC, Contag C, Crawford JM, Wang K, Reisdorph B, Appelman H, Turgeon DK, Meyer C, Wang T. Seeing it through: translational validation of new medical imaging modalities. Biomedical Optics Express. 2012, 3 (4): 764-776.
Liedtke AJ, Crews BC, Daniel CM, Blobaum AL, Kingsley PJ, Ghebreselasie K, Marnett LJ. Cyclooxygenase-1-Selective Inhibitors Based on the (E)-2 '-Des-methyl-sulindac Sulfide Scaffold. Journal of Medicinal Chemistry. 2012, 55 (5): 2287-2300.
Akingbade D, Kingsley PJ, Shuck SC, Cooper T, Carnahan R, Szekely J. Selection of Monoclonal Antibodies Against 6-oxo-M(1)dG and Their Use in an LC-MS/MS Assay for the Presence of 6-oxo-M(1)dG in Vivo. Chemical Research in Toxicology.2012, 25 (2): 454-461.
Brogan JT, Stoops SL, Crews BC, Marnett LJ, Lindsley CW. Total Synthesis of (+)-7-Bromotrypargine and Unnatural Analogues: Biological Evaluation Uncovers Activity at CNS Targets of Therapeutic Relevance. ACS Chemical Neuroscience. 2011, 2 (11): 633-639.