Resistance of human pathogens to anti-infective agents poses a serious threat to human health and requires sustained efforts to develop new therapies. The essential methylerythritol phosphate (MEP) pathway for isoprenoid biosynthesis is widespread in human pathogens, including some of the most deadly infections caused by M. tuberculosis and P. falciparum. Several of the enzymes in this pathway catalyze unprecedented reactions, making them particularly attractive as targets for the development of selective inhibitors. Our long term goal is to understand catalysis in the MEP pathway toward the development of inhibitors targeting isoprenoid biosynthesis in pathogens. This proposal describes studies to examine three intriguing MEP pathway enzymes, IspG, DXP synthase and IspF.
Specific Aims 1 and 3 focus on mechanistic studies and inhibitor development of the enzymes that generate and utilize cyclodiphosphate intermediate, methylerythritol cyclodiphosphate (MEcPP). The goal of Specific Aim 2 is to understand catalysis of DXP synthase in C-N bond formation to generate the medicinally useful hydroxamic acid compound class. Anti-infective agents developed to target these enzymes have the potential to broadly impact the treatment of deadly infectious diseases.

Public Health Relevance

Drug resistance in human pathogens is a global health concern requiring sustained efforts to develop new strategies for treatment of life threatening infections. The proposed studies examine the mechanistically intriguing essential methylerythritol phosphate pathway enzymes which are widespread in human pathogens. Mechanistic studies of these enzymes will lead to the development of new anti-infective agents.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM084998-03
Application #
8269734
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Anderson, Vernon
Project Start
2010-08-01
Project End
2015-05-31
Budget Start
2012-06-01
Budget End
2013-05-31
Support Year
3
Fiscal Year
2012
Total Cost
$316,602
Indirect Cost
$123,552
Name
Johns Hopkins University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Smith, Jessica M; Warrington, Nicole V; Vierling, Ryan J et al. (2014) Targeting DXP synthase in human pathogens: enzyme inhibition and antimicrobial activity of butylacetylphosphonate. J Antibiot (Tokyo) 67:77-83
Hain, Adelaide U P; Bartee, David; Sanders, Natalie G et al. (2014) Identification of an Atg8-Atg3 protein-protein interaction inhibitor from the medicines for Malaria Venture Malaria Box active in blood and liver stage Plasmodium falciparum parasites. J Med Chem 57:4521-31
Afanador, Gustavo A; Matthews, Krista A; Bartee, David et al. (2014) Redox-dependent lipoylation of mitochondrial proteins in Plasmodium falciparum. Mol Microbiol 94:156-71
Brammer Basta, Leighanne A; Patel, Hetalben; Kakalis, Lazaros et al. (2014) Defining critical residues for substrate binding to 1-deoxy-D-xylulose 5-phosphate synthase--active site substitutions stabilize the predecarboxylation intermediate C2?-lactylthiamin diphosphate. FEBS J 281:2820-37
Bitok, J Kipchirchir; Meyers, Caren Freel (2013) Synthesis and evaluation of stable substrate analogs as potential modulators of cyclodiphosphate synthase IspF. Medchemcomm 4:130-134
Morris, Francine; Vierling, Ryan; Boucher, Lauren et al. (2013) DXP synthase-catalyzed C-N bond formation: nitroso substrate specificity studies guide selective inhibitor design. Chembiochem 14:1309-15
Brammer, Leighanne A; Smith, Jessica M; Wade, Herschel et al. (2011) 1-Deoxy-D-xylulose 5-phosphate synthase catalyzes a novel random sequential mechanism. J Biol Chem 286:36522-31