Michael R. Waterman, Ph.D.
Natalie Overall Warren Distinguished Professor and Chair of Biochemistry
Ph.D., University of Oregon, 1969 (Biochemistry)
This laboratory studies structure/function relationships in cytochrome P450 monooxygenases. One enzyme of interest is CYP51 (sterol 14alpha-demethylase), which is required for sterol biosynthesis in different biological kingdoms and is the most widely distributed member of the CYP51 superfamily of genes. The x-ray structure of CYP51 has been determined from Mycobacterium tuberculosis to 2.1 Å and is being used to assist in answering three fundamental questions about CYP51: 1) What is the minimal amino acid requirement for CYP51? 2) What are the amino acids in CYP51 required for substrate specificity? 3) What is the structural basis of azole resistance in the Candida albicans CYP51? Answers to these questions will provide detailed understanding of CYP51 structure/function. A second system under investigation is the group of eighteen cytochrome P450s in Streptomyces coelicolor. Different strains of this soil bacterium produce 75% of commercially available antibiotics. It is believed that several of these P450s are involved in antibiotic biosynthesis while others detoxify chemicals from the environment. The characterization of this group of P450s may facilitate the design of modified enzymes, producing new and more potent antibiotics. The x-ray structure of CYP154C1 was recently solved from S. coelicolor, which can participate in vitro in antibiotic biosynthesis as the beginning step in characterizing these P450s. Structures for CYP154A1 and 158A2 have also been determined. Modified forms of S. coelicolor may also be used for bioremediation of xenobiotic contaminants in the environment.
Recent Publications
Zhao B, Lamb DC, Lei L, Kelly SL, Yuan H, Hachey DL, Waterman MR. Different binding modes of two flaviolin substrate molecules in cytochrome P450 158A1 (CYP158A1) compared to CYP158A2. Biochemistry. 2007 46:8725-8733.
Chun YJ, Shimada T, Sanchez-Ponce R, Martin MV, Lei L, Zhao B, Kelly SL, Waterman MR, Lamb DC, Guengerich FP. Electron transport pathway for a streptomyces cytochrome P450: cytochrome P450 105D5-catalyzed fatty acid hydroxylation in Streptomyces coelicolor A3(2). J Biol Chem. 2007 282:17486-500.
Rupasinghe S, Schuler MA, Kagawa N, Yuan H, Lei L, Zhao B, Kelly SL, Waterman MR, Lamb DC. The cytochrome P450 gene family CYP157 does not contain EXXR in the K-helix reducing the absolute conserved P450 residues to a single cysteine. FEBS Lett. 2006 580:6338-42.
Chun YJ, Shimada T, Waterman MR, Guengerich FP. Understanding electron transport systems of Streptomyces cytochrome P450. Biochem Soc Trans. 2006 34:1183-5.
Lepesheva GI, Hargrove TY, Ott RD, Nes WD, Waterman MR. Biodiversity of CYP51 in trypanosomes. Biochem Soc Trans. 2006 34:1161-4.
Lepesheva GI, Waterman MR. Sterol 14alpha-demethylase cytochrome P450 (CYP51), a P450 in all biological kingdoms. Biochim Biophys Acta. 2007 1770:467-77. Review.
Lamb DC, Guengerich FP, Kelly SL, Waterman MR. Exploiting Streptomyces coelicolor A3(2) P450s as a model for application in drug discovery. Expert Opin Drug Metab Toxicol. 2006 2:27-40.
Sherman DH, Li S, Yermalitskaya LV, Kim Y, Smith JA, Waterman MR, Podust LM. The structural basis for substrate anchoring, active site selectivity, and product formation by P450 PikC from Streptomyces venezuelae. J Biol Chem. 2006 281:26289-97.
McLean KJ, Warman AJ, Seward HE, Marshall KR, Girvan HM, Cheesman MR, Waterman MR, Munro AW. Biophysical characterization of the sterol demethylase P450 from Mycobacterium tuberculosis, its cognate ferredoxin, and their interactions. Biochemistry. 2006 45:8427-43.
Zhao B, Waterman MR, Isin EM, Sundaramoorthy M, Podust LM. Ligand-assisted inhibition in cytochrome P450 158A2 from Streptomyces coelicolor A3(2). Biochemistry. 2006 45:7493-500.
Nakagawa K, Holla VR, Wei Y, Wang WH, Gatica A, Wei S, Mei S, Miller CM, Cha DR, Price E Jr, Zent R, Pozzi A, Breyer MD, Guan Y, Falck JR, Waterman MR, Capdevila JH. Salt-sensitive hypertension is associated with dysfunctional Cyp4a10 gene and kidney epithelial sodium channel. J Clin Invest. 2006 116:1696-702.
Tosha T, Kagawa N, Ohta T, Yoshioka S, Waterman MR, Kitagawa T. Raman evidence for specific substrate-induced structural changes in the heme pocket of human cytochrome P450 aromatase during the three consecutive oxygen activation steps. Biochemistry. 2006 45:5631-40.
Arase M, Waterman MR, Kagawa N. Purification and characterization of bovine steroid 21-hydroxylase (P450c21) efficiently expressed in Escherichia coli. Biochem Biophys Res Commun. 2006 344:400-5.
Zhou W, Lepesheva GI, Waterman MR, Nes WD. Mechanistic analysis of a multiple product sterol methyltransferase implicated in ergosterol biosynthesis in Trypanosoma brucei. J Biol Chem. 2006 281:6290-6.
Lamb DC, Kim Y, Yermalitskaya LV, Yermalitsky VN, Lepesheva GI, Kelly SL, Waterman MR, Podust LM. A second FMN binding site in yeast NADPH-cytochrome P450 reductase suggests a mechanism of electron transfer by diflavin reductases. Structure. 2006 14:51-61.
Lepesheva GI, Zaitseva NG, Nes WD, Zhou W, Arase M, Liu J, Hill GC, Waterman MR. CYP51 from Trypanosoma cruzi: a phyla-specific residue in the B' helix defines substrate preferences of sterol 14alpha-demethylase. J Biol Chem. 2006 281:3577-85.