Journal article
Structural Characterization of the Reaction and Substrate Specificity Mechanisms of Pathogenic Fungal Acetyl-CoA Synthetases
ACS chemical biology, Vol.16(8), pp.1587-1599
08/20/2021
DOI: 10.1021/acschembio.1c00484
PMCID: PMC8383264
PMID: 34369755
Appears in UI Libraries Support Open Access
Abstract
Acetyl CoA synthetases (ACSs) are Acyl-CoA/NRPS/Luciferase (ANL) superfamily enzymes that couple acetate with CoA to generate acetyl CoA, a key component of central carbon metabolism in eukaryotes and prokaryotes. Normal mammalian cells are not dependent on ACSs, while tumor cells, fungi, and parasites rely on acetate as a precursor for acetyl CoA. Consequently, ACSs have emerged as a potential drug target. As part of a program to develop antifungal ACS inhibitors, we characterized fungal ACSs from five diverse human fungal pathogens using biochemical and structural studies. ACSs catalyze a two-step reaction involving adenylation of acetate followed by thioesterification with CoA. Our structural studies captured each step of these two half-reactions including the acetyl-adenylate intermediate of the first half-reaction in both the adenylation conformation and the thioesterification conformation and thus provide a detailed picture of the reaction mechanism. We also used a systematic series of increasingly larger alkyl adenosine esters as chemical probes to characterize the structural basis of the exquisite ACS specificity for acetate over larger carboxylic acid substrates. Consistent with previous biochemical and genetic data for other enzymes, structures of fungal ACSs with these probes bound show that a key tryptophan residue limits the size of the alkyl binding site and forces larger alkyl chains to adopt high energy conformers, disfavoring their efficient binding. Together, our analysis provides highly detailed structural models for both the reaction mechanism and substrate specificity that should be useful in designing selective inhibitors of eukaryotic ACSs as potential anticancer, antifungal, and antiparasitic drugs.
Details
- Title: Subtitle
- Structural Characterization of the Reaction and Substrate Specificity Mechanisms of Pathogenic Fungal Acetyl-CoA Synthetases
- Creators
- Andrew J Jezewski - Roy J. and Lucille A. Carver College of MedicineKaty M Alden - Roy J. and Lucille A. Carver College of MedicineTaiwo E Esan - Northern Illinois UniversityNicholas D DeBouver - UCB Pharma (Belgium)Jan Abendroth - UCB Pharma (Belgium)Jameson C Bullen - UCB Pharma (Belgium)Brandy M Calhoun - UCB Pharma (Belgium)Kristy T Potts - UCB Pharma (Belgium)Daniel M Murante - Roy J. and Lucille A. Carver College of MedicineTimothy J Hagen - Northern Illinois UniversityDavid Fox - UCB Pharma (Belgium)Damian J Krysan - Roy J. and Lucille A. Carver College of Medicine
- Resource Type
- Journal article
- Publication Details
- ACS chemical biology, Vol.16(8), pp.1587-1599
- DOI
- 10.1021/acschembio.1c00484
- PMID
- 34369755
- PMCID
- PMC8383264
- NLM abbreviation
- ACS Chem Biol
- ISSN
- 1554-8929
- eISSN
- 1554-8937
- Publisher
- American Chemical Society
- Number of pages
- 13
- Grant note
- 085P1000817 / Michigan Technology Tri-Corridor Canadian Institutes of Health Research (CIHR); Canadian Institutes of Health Research (CIHR) HHSN272201200025C; HHSN272200700057C / NIAID; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Allergy & Infectious Diseases (NIAID) HHSN272201700059C / National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Department of Health and Human Services Northern Illinois University's Molecular Analysis Core Shimadzu Scientific Instruments University of Saskatchewan Government of Saskatchewan Canada Foundation for Innovation (CFI); Canada Foundation for Innovation DE-AC02-06CH11357 / DOE Office of Science; United States Department of Energy (DOE) National Research Council (NRC) Michigan Economic Development Corporation Natural Sciences and Engineering Research Council (NSERC); Natural Sciences and Engineering Research Council of Canada (NSERC)
- Language
- English
- Date published
- 08/20/2021
- Academic Unit
- Microbiology and Immunology; Stead Family Department of Pediatrics; Infectious Disease (Pediatrics)
- Record Identifier
- 9984230429202771
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