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Insights into dynamic changes in ADC-7 and P99 cephalosporinases using small angle x-ray scattering (SAXS)

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title Insights into dynamic changes in ADC-7 and P99 cephalosporinases using small angle x-ray scattering (SAXS)
 
Creator Kumar, Ish
Sagar, Amin
Dhiman, Kanika
Bethel , Christopher R
Hujer , Andrea M
Carifi , Justin
., Ashish
Bonomo , Robert A
 
Subject QR Microbiology
 
Description To counter the emergence of β-lactamase (BL) mediated resistance, design of new β-lactamase inhibitors (BLIs) is critical. Many high-resolution crystallographic structures of BL complexed with BLIs are available. However, their impact on BLI design is struggling to keep pace with novel and emerging variants. Small angle x-ray scattering (SAXS) in combination with molecular modeling is a useful tool to determine dynamic structures of macromolecules in solution. An important application of SAXS is to determine the conformational changes that occur when BLI bind to BL. To probe if conformational dynamics occur in class C cephalosporinases, we studied SAXS profiles of two clinically relevant class C β-lactamases, Acinetobacter baumannii ADC-7 and Enterobacter cloacae P99 in apo format complexed with BLIs. Importantly, SAXS data analysis demonstrated that in solution, these representative class C enzymes remain monomeric and did not show the associated assemblies that were seen in various crystal structures. SAXS data acquired for ADC-7 and P99, in apo and inhibitor bound states, clearly showed that these enzymes undergo detectable conformational changes, and these class C β-lactamases also close upon binding inhibitors as does BlaC. Further analysis revealed that addition of inhibitor led to the compacting of a range of residues around the active site, indicating that the conformational changes that both P99 and ADC-7 undergo are central to inhibitor recognition and efficacy. Our findings support the importance of exploring conformational changes using SAXS analysis in the design of future BLIs.
 
Publisher Adenine Press
 
Date 2023-08
 
Type Article
PeerReviewed
 
Relation https://www.tandfonline.com/doi/full/10.1080/07391102.2023.2240427
http://crdd.osdd.net/open/3186/
 
Identifier Kumar, Ish and Sagar, Amin and Dhiman, Kanika and Bethel , Christopher R and Hujer , Andrea M and Carifi , Justin and ., Ashish and Bonomo , Robert A (2023) Insights into dynamic changes in ADC-7 and P99 cephalosporinases using small angle x-ray scattering (SAXS). Journal of Biomolecular Structure and Dynamics, 14. pp. 1-13. ISSN 0739-1102