Structural and Functional Characterization of Methicillin-Resistant Staphylococcus aureus’s Class IIb Fructose 1,6-Bisphosphate Aldolase

Capodagli, Glenn C.; Lee, Stephen A.; Boehm, Kyle J.; Brady, Kristin M.; Pegan, Scott D.

doi:10.1021/bi501141t

Title: Structural and Functional Characterization of Methicillin-Resistant Staphylococcus aureus’s Class IIb Fructose 1,6-Bisphosphate Aldolase

Full Record
Other Related Research

Abstract

Staphylococcus aureus is one of the most common nosocomial sources of soft-tissue and skin infections and has more recently become prevalent in the community setting as well. Since the use of penicillins to combat S. aureus infections in the 1940s, the bacterium has been notorious for developing resistances to antibiotics, such as methicillin-resistant Staphylococcus aureus (MRSA). With the persistence of MRSA as well as many other drug resistant bacteria and parasites, there is a growing need to focus on new pharmacological targets. Recently, class II fructose 1,6-bisphosphate aldolases (FBAs) have garnered attention to fill this role. Regrettably, scarce biochemical data and no structural data are currently available for the class II FBA found in MRSA (SaFBA). With the recent finding of a flexible active site zinc-binding loop (Z-Loop) in class IIa FBAs and its potential for broad spectrum class II FBA inhibition, the lack of information regarding this feature of class IIb FBAs, such as SaFBA, has been limiting for further Z-loop inhibitor development. Therefore, we elucidated the crystal structure of SaFBA to 2.1 Å allowing for a more direct structural analysis of SaFBA. Furthermore, we determined the KM for one of SaFBA’s substrates, fructose 1,6-bisphosphate, as well as performedmore »« less

Authors:

Capodagli, Glenn C. ^[1]; Lee, Stephen A. ^[1]; Boehm, Kyle J. ^[1]; Brady, Kristin M. ^[1]; Pegan, Scott D. ^[2]

Department of Chemistry and Biochemistry, University of Denver, Denver, Colorado 80208, United States
Department of Pharmaceutical and Biomedical Sciences, University of Georgia, Athens, Georgia 30602, United States

Publication Date:: Fri Nov 21 00:00:00 EST 2014

Research Org.:: Univ. of Denver, Denver, CO (United States); Univ. of Georgia, Athens, GA (United States)

Sponsoring Org.:: USDOE Advanced Research Projects Agency - Energy (ARPA-E); USDOE Office of Science (SC), Basic Energy Sciences (BES)

OSTI Identifier:: 1164331

Alternate Identifier(s):: OSTI ID: 1345591

Grant/Contract Number:: AC02-05CH11231

Resource Type:: Published Article

Journal Name:: Biochemistry

Additional Journal Information:: Journal Name: Biochemistry Journal Volume: 53 Journal Issue: 48; Journal ID: ISSN 0006-2960

Publisher:: American Chemical Society

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES

Citation Formats


                    Capodagli, Glenn C., Lee, Stephen A., Boehm, Kyle J., Brady, Kristin M., and Pegan, Scott D. Structural and Functional Characterization of Methicillin-Resistant Staphylococcus aureus’s Class IIb Fructose 1,6-Bisphosphate Aldolase.  United States: N. p., 2014. 
Web.  doi:10.1021/bi501141t.

Copy to clipboard


                    Capodagli, Glenn C., Lee, Stephen A., Boehm, Kyle J., Brady, Kristin M., & Pegan, Scott D. Structural and Functional Characterization of Methicillin-Resistant Staphylococcus aureus’s Class IIb Fructose 1,6-Bisphosphate Aldolase.  United States.  https://doi.org/10.1021/bi501141t

Copy to clipboard


                    Capodagli, Glenn C., Lee, Stephen A., Boehm, Kyle J., Brady, Kristin M., and Pegan, Scott D. Fri .  
"Structural and Functional Characterization of Methicillin-Resistant Staphylococcus aureus’s Class IIb Fructose 1,6-Bisphosphate Aldolase".  United States.  https://doi.org/10.1021/bi501141t.

Copy to clipboard


                    
@article{osti_1164331,

  title        = {Structural and Functional Characterization of Methicillin-Resistant Staphylococcus aureus’s Class IIb Fructose 1,6-Bisphosphate Aldolase},

  author       = {Capodagli, Glenn C. and Lee, Stephen A. and Boehm, Kyle J. and Brady, Kristin M. and Pegan, Scott D.},

  abstractNote = {Staphylococcus aureus is one of the most common nosocomial sources of soft-tissue and skin infections and has more recently become prevalent in the community setting as well. Since the use of penicillins to combat S. aureus infections in the 1940s, the bacterium has been notorious for developing resistances to antibiotics, such as methicillin-resistant Staphylococcus aureus (MRSA). With the persistence of MRSA as well as many other drug resistant bacteria and parasites, there is a growing need to focus on new pharmacological targets. Recently, class II fructose 1,6-bisphosphate aldolases (FBAs) have garnered attention to fill this role. Regrettably, scarce biochemical data and no structural data are currently available for the class II FBA found in MRSA (SaFBA). With the recent finding of a flexible active site zinc-binding loop (Z-Loop) in class IIa FBAs and its potential for broad spectrum class II FBA inhibition, the lack of information regarding this feature of class IIb FBAs, such as SaFBA, has been limiting for further Z-loop inhibitor development. Therefore, we elucidated the crystal structure of SaFBA to 2.1 Å allowing for a more direct structural analysis of SaFBA. Furthermore, we determined the KM for one of SaFBA’s substrates, fructose 1,6-bisphosphate, as well as performed mode of inhibition studies for an inhibitor that takes advantage of the Z-loop’s flexibility. Altogether the data offers insight into a class IIb FBA from a pervasively drug resistant bacterium and a comparison of Z-loops and other features between the different subtypes of class II FBAs.},

  doi          = {10.1021/bi501141t},

  journal      = {Biochemistry},

  number       = 48,

  volume       = 53,

  place        = {United States},

  year         = {Fri Nov 21 00:00:00 EST 2014},

  month        = {Fri Nov 21 00:00:00 EST 2014}

}

Copy to clipboard

Journal Article:

Free Publicly Available Full Text

Publisher's Version of Record
https://doi.org/10.1021/bi501141t

Other availability

Search WorldCat to find libraries that may hold this journal

Citation Metrics:

Cited by: 13 works

Citation information provided by
Web of Science

Save / Share:

Export Metadata

Save to My Library

Similar Records in DOE PAGES and OSTI.GOV collections:

Structural Basis for Catalysis of a Tetrameric Class IIa Fructose 1,6-Bisphosphate Aldolase from Mycobacterium tuberculosis

Journal Article Pegan, Scott D ; Ruskseree, Kamolchanok ; Franzblau, Scott G ; ... - J. Mol. Biol.

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), currently infects one-third of the world's population in its latent form. The emergence of multidrug-resistant and extensive drug-resistant strains has highlighted the need for new pharmacological targets within M. tuberculosis. The class IIa fructose 1,6-bisphosphate aldolase (FBA) enzyme from M. tuberculosis (MtFBA) has been proposed as one such target since it is upregulated in latent TB. Since the structure of MtFBA has not been determined and there is little information available on its reaction mechanism, we sought to determine the X-ray structure of MtFBA in complex with its substrates. By lowering themore »« less
https://doi.org/10.1016/j.jmb.2009.01.003
Active Site Loop Dynamics of a Class IIa Fructose 1,6-Bisphosphate Aldolase from Mycobacterium tuberculosis

Journal Article Pegan, Scott D. ; Rukseree, Kamolchanok ; Capodagli, Glenn C. ; ... - Biochemistry

The class II fructose 1,6-bisphosphate aldolases (FBAs, EC 4.1.2.13) comprises one of two families of aldolases. Instead of forming a Schiff base intermediate using an ε-amino group of a lysine side chain, class II FBAs utilize Zn(II) to stabilize a proposed hydroxyenolate intermediate (HEI) in the reversible cleavage of fructose 1,6-bisphosphate, forming glyceraldehyde 3-phosphate and dihydroxyacetone phosphate (DHAP). As class II FBAs have been shown to be essential in pathogenic bacteria, focus has been placed on these enzymes as potential antibacterial targets. Although structural studies of class II FBAs from Mycobacterium tuberculosis (MtFBA), other bacteria, and protozoa have been reported,more »« less
https://doi.org/10.1021/bi300928u
Crystal Structures of Wild-type and Mutant Methicillin-resistant Staphylococcus aureus Dihydrofolate Reductase Reveal an Alternative Conformation of NADPH that may be Linked to Trimethoprim Resistance

Journal Article Frey, K ; Liu, J ; Lombardo, M ; ... - Journal of Molecular Biology

Both hospital- and community-acquired Staphylococcus aureus infections have become major health concerns in terms of morbidity, suffering and cost. Trimethoprim-sulfamethoxazole (TMP-SMZ) is an alternative treatment for methicillin-resistant S. aureus (MRSA) infections. However, TMP-resistant strains have arisen with point mutations in dihydrofolate reductase (DHFR), the target for TMP. A single point mutation, F98Y, has been shown biochemically to confer the majority of this resistance to TMP. Using a structure-based approach, we have designed a series of novel propargyl-linked DHFR inhibitors that are active against several trimethoprim-resistant enzymes. We screened this series against wild-type and mutant (F98Y) S. aureus DHFR and foundmore »« less
https://doi.org/10.1016/j.jmb.2009.02.045
Structural comparison of chromosomal and exogenous dihydrofolate reductase from Staphylococcus aureus in complex with the potent inhibitor trimethoprim

Journal Article Heaslet, Holly ; Harris, Melissa ; Fahnoe, Kelly ; ... - Proteins

Dihydrofolate reductase (DHFR) is the enzyme responsible for the NADPH-dependent reduction of 5,6-dihydrofolate to 5,6,7,8-tetrahydrofolate, an essential cofactor in the synthesis of purines, thymidylate, methionine, and other key metabolites. Because of its importance in multiple cellular functions, DHFR has been the subject of much research targeting the enzyme with anticancer, antibacterial, and antimicrobial agents. Clinically used compounds targeting DHFR include methotrexate for the treatment of cancer and diaminopyrimidines (DAPs) such as trimethoprim (TMP) for the treatment of bacterial infections. DAP inhibitors of DHFR have been used clinically for >30 years and resistance to these agents has become widespread. Methicillin-resistant Staphylococcusmore »« less
https://doi.org/10.1002/prot.22383
Feedback-Based, System-Level Properties of Vertebrate-Microbial Interactions

Journal Article Rivas, Ariel L. ; Jankowski, Mark D. ; Piccinini, Renata ; ... - PLoS ONE

Improved characterization of infectious disease dynamics is required. To that end, three-dimensional (3D) data analysis of feedback-like processes may be considered. To detect infectious disease data patterns, a systems biology (SB) and evolutionary biology (EB) approach was evaluated, which utilizes leukocyte data structures designed to diminish data variability and enhance discrimination. Using data collected from one avian and two mammalian (human and bovine) species infected with viral, parasite, or bacterial agents (both sensitive and resistant to antimicrobials), four data structures were explored: (i) counts or percentages of a single leukocyte type, such as lymphocytes, neutrophils, or macrophages (the classic approach),more »« less
https://doi.org/10.1371/journal.pone.0053984

Full Text Available

Similar Records