skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Dissecting the Structural Elements for the Activation of β-Ketoacyl-(Acyl Carrier Protein) Reductase from Vibrio cholerae

Journal Article · · Journal of Bacteriology
DOI:https://doi.org/10.1128/JB.00360-15· OSTI ID:1239405
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2];  [1]
  1. Univ. of Virginia, Charlottesville, VA (United States)
  2. J. Craig Venter Inst., Rockville, MD (United States)
+ Show Author Affiliations

β-Ketoacyl-(acyl carrier protein) reductase (FabG) catalyzes the key reductive reaction in the elongation cycle of fatty acid synthesis (FAS), which is a vital metabolic pathway in bacteria and a promising target for new antibiotic development. The activation of the enzyme is usually linked to the formation of a catalytic triad and cofactor binding, and crystal structures of FabG from different organisms have been captured in either the active or inactive conformation. However, the structural elements which enable activation of FabG require further exploration. Here we report the findings of structural, enzymatic, and binding studies of the FabG protein found in the causative agent of cholera, Vibrio cholerae (vcFabG). vcFabG exists predominantly as a dimer in solution and is able to self-associate to form tetramers, which is the state seen in the crystal structure. The formation of the tetramer may be promoted by the presence of the cofactor NADP(H). The transition between the dimeric and tetrameric states of vcFabG is related to changes in the conformations of the α4/α5 helices on the dimer-dimer interface. Two glycine residues adjacent to the dimer interface (G92 and G141) are identified to be the hinge for the conformational changes, while the catalytic tyrosine (Y155) and a glutamine residue that forms hydrogen bonds to both loop β4-α4 and loop β5-α5 (Q152) stabilize the active conformation. The functions of the aforementioned residues were confirmed by binding and enzymatic assays for the corresponding mutants. This study describes the results of structural, enzymatic, and binding studies of FabG from Vibrio cholerae (vcFabG). In this work, we dissected the structural elements responsible for the activation of vcFabG. The structural information provided here is essential for the development of antibiotics specifically targeting bacterial FabG, especially for the multidrug-resistant strains of V. cholerae.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER); NIAID; National Inst. of Allergy and Infectious Diseases; National Inst. of Health; U.S. Dept. of Health and Human Services
Grant/Contract Number:
AC02-06CH11357; HHSN272200700058C; HHSN272201200026C
OSTI ID:
1239405
Journal Information:
Journal of Bacteriology, Vol. 198, Issue 3; ISSN 0021-9193
Publisher:
American Society for MicrobiologyCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 12 works
Citation information provided by
Web of Science

References (54)

Multimeric Options for the Auto-Activation of the Saccharomyces cerevisiae FAS Type I Megasynthase journal August 2009
Automated protein model building combined with iterative structure refinement
  • Perrakis, Anastassis; Morris, Richard; Lamzin, Victor S.
  • Nature Structural Biology, Vol. 6, Issue 5, p. 458-463 https://doi.org/10.1038/8263
journal May 1999
Short-chain dehydrogenases/reductases (SDRs): Coenzyme-based functional assignments in completed genomes journal September 2002
The kinetics of enzyme-catalyzed reactions with two or more substrates or products journal January 1963
Overview of the CCP 4 suite and current developments journal March 2011
Crystal structure and fluorescence studies reveal the role of helical dimeric interface of staphylococcal fabg1 in positive cooperativity for NADPH journal February 2012
Fluorescence-Energy Transfer in Human Estradiol 17beta-Dehydrogenase-NADPH Complex and Studies on the Coenzyme Binding journal January 1996
Crystal Structure and Enzyme Kinetics of the ( S )-Specific 1-Phenylethanol Dehydrogenase of the Denitrifying Bacterium Strain EbN1 † , ‡ journal January 2006
The Structural Biology of type ii Fatty acid Biosynthesis journal June 2005
Crystal Structure of MabA from Mycobacterium tuberculosis, a Reductase involved in Long-chain Fatty Acid Biosynthesis journal July 2002
The Structural Biology Center 19ID undulator beamline: facility specifications and protein crystallographic results journal December 2005
Study of protein subunit association equilibria by elution gel chromatography book January 1979
Comparative evaluation of antimicrobials for textile applications journal March 2013
Gapped BLAST and PSI-BLAST: a new generation of protein database search programs journal September 1997
Cofactor-Induced Conformational Rearrangements Establish a Catalytically Competent Active Site and a Proton Relay Conduit in FabG journal March 2004
Improved methods for building protein models in electron density maps and the location of errors in these models journal March 1991
Inhibitory effects on bacterial growth and b-ketoacyl-ACP reductase by different species of maple leaf extracts and tannic acid journal May 2009
3-Oxoacyl-[ACP] reductase from oilseed rape (Brassica napus) journal April 1992
Transcriptional Analysis of Essential Genes of theEscherichia coli Fatty Acid Biosynthesis Gene Cluster by Functional Replacement with the Analogous Salmonella typhimurium Gene Cluster journal July 1998
Fatty acid synthase as a potential therapeutic target in cancer journal April 2010
Inference of Macromolecular Assemblies from Crystalline State journal September 2007
Structural insight into the catalytic mechanism of gluconate 5-dehydrogenase from Streptococcus suis : Crystal structures of the substrate-free and quaternary complex enzymes journal February 2009
Cholera: a great global concern journal July 2011
Mycobacterium tuberculosis β-ketoacyl-ACP reductase: α-Secondary kinetic isotope effects and kinetic and equilibrium mechanisms of substrate binding journal March 2008
Improvement of macromolecular electron-density maps by the simultaneous application of real and reciprocal space constraints journal January 1993
Crystal structure and function of the isoniazid target of Mycobacterium tuberculosis journal March 1995
A short history of SHELX journal December 2007
Crystallization of the NADP-dependent β-keto acyl-carrier protein reductase from Brassica napus journal January 2000
HKL -3000: the integration of data reduction and structure solution – from diffraction images to an initial model in minutes journal July 2006
Structure of β-Ketoacyl-[acyl carrier protein] Reductase from Escherichia coli :  Negative Cooperativity and Its Structural Basis † , ‡ journal October 2001
Coot model-building tools for molecular graphics journal November 2004
New Insight into the Mechanism of Action of and Resistance to Isoniazid:  Interaction of Mycobacterium tuberculosis enoyl-ACP Reductase with INH-NADP journal August 2007
Structural and Functional Studies on SCO1815:  A β-Ketoacyl-Acyl Carrier Protein Reductase from Streptomyces coelicolor A3(2) ∞ journal November 2006
REFMAC 5 for the refinement of macromolecular crystal structures journal March 2011
Structure validation by Cα geometry: ϕ,ψ and Cβ deviation journal January 2003
Expression and Purification of Soluble His6-Tagged TEV Protease book January 2009
Mycobacterium tuberculosis β-Ketoacyl-Acyl Carrier Protein (ACP) Reductase:  Kinetic and Chemical Mechanisms † journal October 2006
Crystal structure of FabG4 from Mycobacterium tuberculosis reveals the importance of C-terminal residues in ketoreductase activity journal April 2011
MabA (FabG1), a Mycobacterium tuberculosis protein involved in the long-chain fatty acid elongation system FAS-II journal April 2002
The Crystal Structure of a Mammalian Fatty Acid Synthase journal September 2008
Dali: a network tool for protein structure comparison journal November 1995
A New Vector for High-Throughput, Ligation-Independent Cloning Encoding a Tobacco Etch Virus Protease Cleavage Site journal June 2002
Ligand-induced fit in mycobacterial MabA: The sequence-specific C-terminus locks the conformational change
  • Cohen-Gonsaud, Martin; Ducasse-Cabanot, Stéphanie; Quemard, Annaïk
  • Proteins: Structure, Function, and Bioinformatics, Vol. 60, Issue 3 https://doi.org/10.1002/prot.20494
journal June 2005
Analyses of co-operative transitions in Plasmodium falciparumβ-ketoacyl acyl carrier protein reductase upon co-factor and acyl carrier protein binding: Plasmodium falciparumβ-ketoacyl ACP reductase journal August 2006
Discovery of an Allosteric Inhibitor Binding Site in 3-Oxo-acyl-ACP Reductase from Pseudomonas aeruginosa journal September 2013
Kinetic, inhibition and structural studies on 3-oxoacyl-ACP reductase from Plasmodium falciparum, a key enzyme in fatty acid biosynthesis journal December 2005
Automated structure solution, density modification and model building journal October 2002
ConSurf 2010: calculating evolutionary conservation in sequence and structure of proteins and nucleic acids journal May 2010
Inhibition of β-Ketoacyl-Acyl Carrier Protein Synthases by Thiolactomycin and Cerulenin: STRUCTURE AND MECHANISM journal October 2000
T-Coffee: a web server for the multiple sequence alignment of protein and RNA sequences using structural information and homology extension journal May 2011
Structure of zinc-independent sorbitol dehydrogenase from Rhodobacter sphaeroides at 2.4 Å resolution journal March 2005
[20] Processing of X-ray diffraction data collected in oscillation mode book January 1997
Is bacterial fatty acid synthesis a valid target for antibacterial drug discovery? journal October 2011
Crystal structure of a 3-oxoacyl-(acylcarrier protein) reductase (BA3989) from Bacillus anthracis at 2.4-Å resolution journal September 2007

Cited By (1)

Structural rearrangements occurring upon cofactor binding in the Mycobacterium smegmatis β-ketoacyl-acyl carrier protein reductase MabA journal April 2018

Similar Records

X-ray structures of uridine phosphorylase from Vibrio cholerae in complexes with uridine, thymidine, uracil, thymine, and phosphate anion: Substrate specificity of bacterial uridine phosphorylases
Journal Article · Tue Nov 15 00:00:00 EST 2016 · Crystallography Reports · OSTI ID:1239405
+4 more
Conserved residue His-257 of Vibrio cholerae flavin transferase ApbE plays a critical role in substrate binding and catalysis
Journal Article · Fri Jul 26 00:00:00 EDT 2019 · Journal of Biological Chemistry · OSTI ID:1239405
+10 more
The crystal structure of Vibrio cholerae (6-4) photolyase reveals interactions with cofactors and a DNA-binding region
Journal Article · Wed Dec 14 00:00:00 EST 2022 · Journal of Biological Chemistry · OSTI ID:1239405

Related Subjects

59 BASIC BIOLOGICAL SCIENCES