Steven L. Roderick, Ph.D.

Structure and Function of Enzymes

In particular, we are interested in bacterial enzymes that are absent from humans which may represent attractive drug targets for agents directed against gram-negative pathogens. One such enzyme is involved in the biosynthesis of lipopolysaccharide (LPS), the predominant lipid present in the outermost membrane monolayer of gram-negative bacteria such as E. coli. The crystal structure of the first enzyme of the LPS biosynthetic pathway, UDP-GlcNAc O-Acyltransferase (LpxA), has allowed us to identify a novel coiled structural domain that we termed a left-handed parallel beta helix (Science 270,997-1000). This domain is characteristic of the hexapeptide acyltransferase superfamily of enzymes of which LpxA is a member.

The laboratory has now solved the crystal structure of seven members of this family with the aim of investigating their catalytic mechanisms of action and developing their potential as antibacterial drug targets. Among the enzymes we are currently studying are a bifunctional acetyltransferase/uridyltransferase capable of synthesizing the UDP-GlcNAc precursor of bacterial cell walls (GlmU), an acetyltransferase responsible for conferring high-level resistance to streptogramin group A antibiotics (VatD), and the key regulatory enzyme of bacterial sulfur assimilation, serine acetyltransferase (SAT).

Publications

Salsi, E. Campanini, B., Bettati, S., Raboni, S., Roderick, S.L., Cook, P.F. and Mozzarelli, A. (2010). "A Two-Step Process Controls the Formation of the Bienzyme Cysteine Synthase Complex." Journal of Biological Chemistry 285, 12813-12822.

Salsi, E., Bayden, A.S., Spyrakis, F., Amadasi, A., Campanini, B., Bettati, S., Dodatko, T., Cozzini, P., Kellogg, G.E., Cook, P.F., Roderick S.L. and Mozzarelli, A. (2010). “Design of O-Acetylserine Sulfhydrylase Inhibitors by Mimicking Nature”. Journal of Medicinal Chemistry 53, 345-356.

Pereira, M.P., Blanchard, J.E., Murphy, C., Roderick, S.L. and Brown, E.D. (2009). “High-Throughput Screening Identifies Novel Inhibitors of the Acetyltransferase Activity of Escherichia coli GlmU.” Antimicrobial Agents and Chemotherapy 53, 2306-2311.

Guan, R., Roderick, S.L., Huang, B. and Cook, P.F. (2008). “Roles of Histidines 154 and 189 and Aspartate 139 in the Active Site of Serine Acetyltransferase from Haemophilus influenzae”. Biochemistry 47, 6322-6328.

Kanno, K., Wu, M.K., Scapa, E.F., Roderick, S.L. and Cohen, D.E. (2007). “Structure and function of phosphatidylcholine transfer protein (PC-TP)/StarD2”. BBA 1771, 654-662.

Olsen, L.R., Vetting, M.W. and Roderick, S.L. (2007). “Structure of the E. coli bifunctional GlmU acetyltransferase active site with substrates and products". Protein Science 16, 1230-1235.

Andreassi, J.L., Bilder, P.W., Vetting, M.W., Roderick, S.L. and Leyh, T.S. (2007). “Crystal structure of the Streptococcus pneumoniae mevalonate kinase in complex with diphosphomevalonate”. Protein Science 16, 983-989.

Pan, H.-J., Agate, D.S., King, B.L., Wu, M.K., Roderick, S.L., Leiter, E.H. and Cohen, D.E. (2006). “A polymorphism in New Zealand inbred mouse strains that inactivates phosphatidylcholine transfer protein”. FEBS Letters 580, 5953-5958.

Vetting, M.W., Hegde, S.S., Fajardo,J.E., Fiser, A., Roderick, S.L., Takiff, H.E. and Blanchard, J.S. (2006). “The Pentapeptide Repeat Proteins”. Biochemistry 45, 1-10.

Campanini, B., Speroni, F., Salsi, E., Cook, P.F., Roderick, S.L., Huang, B., Bettati, S. and Mozzarelli, A. (2005). “Interaction of serine acetyltransferase with O-acetylserine sulfhydrylase active site: Evidence from fluorescence spectroscopy”. Protein Science 14, 2115-2124.

Vetting, M.W., de Carvalho, L.P.S., Roderick, S.L. and Blanchard, J.S. (2005). “A Novel Dimeric Structure of the RimL N^á-acetyltransferase from Salmonella typhimurium”. Journal of Biological Chemistry 280, 22108-22114.

Hegde, S.S., Vetting, M.W., Roderick, S.L., Mitchenall, L.A. , Maxwell, A., Takiff, H.E. and Blanchard, J.S. (2005). “A Fluoroquinolone Resistance Protein from Mycobacterium tuberculosis that Mimics DNA”. Science 308, 1480-1483.

Roderick, S.L. (2005). “The lac operon galactoside acetyltransferase”. Comptes Rendus Biologies 328, 568-575.

Huang, B., Vetting, M.W. and Roderick, S.L. (2005). “The Active Site of O-Acetylserine Sulfhydrylase Is the Anchor Point for Bienzyme Complex Formation with Serine Acetyltransferase”. Journal of Bacteriology 187, 3201-3205.

Johnson, C.M., Roderick, S.L. and Cook, P.F. (2005). “The serine acetyltransferase reaction:  acetyl transfer from an acylpantothenyl donor to an alcohol”. Archives of Biochemistry and Biophysics 433, 85-95.

Vetting, M.W., de Carvalho, L.P.S., Yu, M., Hegde, S.S., Magnet, S., Roderick, S.L. and Blanchard, J.S. (2005). “Structure and functions of the GNAT superfamily of acetyltransferases”. Archives of Biochemistry and Biophysics 433, 212-226.

Johnson, C.M., Huang, B., Roderick, S.L. and Cook, P.F. (2004). “Chemical Mechanism of the Serine Acetyltransferase from Haemophilus influenzae”. Biochemistry 43, 15534-15539.

Johnson, C.M., Huang, B., Roderick, S.L. and Cook, P.F. (2004). "Kinetic mechanism of the serine acetyltransferase from Haemophilus influenzae". Archives of Biochemistry and Biophysics 429, 115-122.

Olsen, L.R., Huang, B., Vetting, M.W. and Roderick, S.L. (2004). "Structure of Serine Acetyltransferase in Complexes with CoA and Its Cysteine Feedback Inhibitor". Biochemistry 43, 6013-6019.

Vetting, M.W., Magnet, S., Nieves, E., Roderick, S.L. and Blanchard, J.S. (2004). "A Bacterial Acetyltransferase Capable of Regioselective N-Acetylation of Antibiotics and Histones". Chemistry and Biology 11, 565-573.

Vetting, M.W., Roderick, S.L., Yu, M. and Blanchard, J.S. (2003). "Crystal structure of mycothiol synthase (Rv0819) from Mycobacterium tuberculosis shows structural homology to the GNAT family of acetyltransferases". Protein Science 12, 1954-1959.

Vetting, M.W., Roderick, S.L., Hedge, S., Magnet, S. and Blanchard, J.S. (2003). "What can structure tell us about in vivo function? The case of aminoglycoside-resistance genes". Biochem Soc Trans 31, 520-522.

Vetting, M.W., Hedge, S.S., Javid-Majd, F., Blanchard, J.S. and Roderick, S.L. (2002). "Structure of aminoglycoside 2'-N-acetyltransferase from Mycobacterium tuberculosis in complex with coenzyme A and aminoglycoside substrates". Nature Structural Biology 9, 653-658.

Roderick, S.L., Chan, W.W., Agate, D.W., Olsen, L.R., Vetting, M.W., Rajashankar, K.R. and Cohen, D.E. (2002). "Structure of human phosphatidylcholine transfer protein in complex with its ligand". Nature Structural Biology 9, 507-511.

Chan, W.W., Roderick, S.L. and Cohen, D.E. (2002). "Human phosphatidylcholine transfer protein: purification, crystallization and preliminary X-ray diffraction". Biochem Biophys Acta 1596, 1-5.

Wang, X.-G., Olsen, L.R. and Roderick, S.L. (2002). "Structure of the lac Operon Galactoside Acetyltransferase". Structure 10, 581-588.

Beaman, T., Vogel, K.W., Drueckhammer, D.G., Blanchard, J.S. and Roderick, S.L. (2002). "Acyl group specificity at the active site of tetrahydrodipicolinate N-succinyltransferase". Protein Science 11, 974-979.

Sugantino, M. and Roderick, S.L. (2002). "Crystal Structure of Vat(D): An Acetyltransferase That Inactivates Streptogramin Group A Antibiotics". Biochemistry 41, 2209-2216.

Olsen, L.R. and Roderick, S.L. (2001). "Structure of the Escherichia coli GlmU Pyrophosphorylase and Acetyltransferase Active Sites". Biochemistry 40, 1913-1929.

Olsen, L.R. and Roderick, S.L. (2001). "Purification, crystallization and preliminary X-ray data for Escherichia coli GlmU: a bifunctional uridyltransferase/acetyltransferase". Acta Crystallographica D57, 296-297.

Feng, L., Chan, W.W., Roderick, S.L. and Cohen, D.E. (2000). "High-Level Expression and Mutagenesis of Recombinant Human Phosphatidylcholine Transfer Protein Using a Synthetic Gene: Evidence for a C-Terminal Membrane Binding Domain". Biochemistry 39, 15399-15409.

Sugantino, M. and Roderick, S.L. (2000). "Expression, Purification and Crystallization of Enterococcus faecium Streptogramin A Acetyltransferase". Acta Crystallographica D56, 640-642.

Wang, X.-G. and Roderick, S.L. (1999). "Expression, Purification, Crystallization and Preliminary X-ray Data of Escherichia coli Galactoside Acetyltransferase". Acta Crystallographica D55, 1955-1957.

Beaman, T.W., Blanchard, J.S. and Roderick, S.L. (1998). "The Active Site Structure and Conformational Change of Tetrahydrodipicolinate N-Succinyltransferase". Biochemistry 37, 10363-10369.

Beaman, T.W., Sugantino, M. and Roderick, S.L. (1998). "Structure of the Hexapeptide Xenobiotic Acetyltransferase from Pseudomonas aeruginosa". Biochemistry 37, 6689-6696.

Tian,Y., Beaman, T.W. and Roderick, S.L. (1997). "Purification and Crystallization of Pseudomonas aeruginosa Chloramphenicol Acetyltransferase". Proteins 28, 298- 300.

Beaman, T.W., Binder, D.A., Blanchard, J.S. and Roderick, S.L. (1997). "Three-Dimensional Structure of Tetrahydrodipicolinate N-Succinyltransferase". Biochemistry 36, 489-494.

Binder, D.A., Blanchard, J.S. and Roderick, S.L. (1996). "Crystallization and Preliminary Crystallographic Analysis of Tetrahydrodipicolinate N-Succinyltransferase". Proteins 26, 115-117.

Raetz, C.R.H. and Roderick, S.L. (1995). "A Left-Handed Parallel Beta Helix in the Structure of UDP-N-Acetylglucosamine Acyltransferase". Science 270, 997-1000.

Pfitzner, U., Raetz, C.R.H. and Roderick, S.L. (1995). "Crystallization of UDP-N-Acetylglucosamine O-Acyltransferase from Escherichia coli". Proteins 22, 191-192.