Klebsiella pneumoniae Carbapenemase-2 (KPC-2), Substitutions at Ambler Position Asp179, and Resistance to Ceftazidime-Avibactam: Unique Antibiotic-Resistant Phenotypes Emerge from β-Lactamase Protein Engineering

MBio
Melissa D BarnesRobert A Bonomo

Abstract

The emergence of Klebsiella pneumoniae carbapenemases (KPCs), β-lactamases that inactivate "last-line" antibiotics such as imipenem, represents a major challenge to contemporary antibiotic therapies. The combination of ceftazidime (CAZ) and avibactam (AVI), a potent β-lactamase inhibitor, represents an attempt to overcome this formidable threat and to restore the efficacy of the antibiotic against Gram-negative bacteria bearing KPCs. CAZ-AVI-resistant clinical strains expressing KPC variants with substitutions in the Ω-loop are emerging. We engineered 19 KPC-2 variants bearing targeted mutations at amino acid residue Ambler position 179 in Escherichia coli and identified a unique antibiotic resistance phenotype. We focus particularly on the CAZ-AVI resistance of the clinically relevant Asp179Asn variant. Although this variant demonstrated less hydrolytic activity, we demonstrated that there was a prolonged period during which an acyl-enzyme intermediate was present. Using mass spectrometry and transient kinetic analysis, we demonstrated that Asp179Asn "traps" β-lactams, preferentially binding β-lactams longer than AVI owing to a decreased rate of deacylation. Molecular dynamics simulations predict that (i) the Asp179Asn variant...Continue Reading

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Citations

Mar 14, 2018·Antimicrobial Agents and Chemotherapy·Melissa D BarnesRobert A Bonomo
Jun 12, 2019·Antimicrobial Agents and Chemotherapy·Melissa D BarnesRobert A Bonomo
Aug 1, 2019·The Journal of Antimicrobial Chemotherapy·Stephan GöttigThomas A Wichelhaus
May 28, 2020·Antimicrobial Agents and Chemotherapy·Laurent PoirelPatrice Nordmann
Oct 24, 2018·The Journal of Antimicrobial Chemotherapy·David M Livermore
May 28, 2019·The Journal of Antimicrobial Chemotherapy·Saoussen OueslatiThierry Naas
Jun 28, 2019·Frontiers in Public Health·Ilias KaraiskosGaryphallia Poulakou
Aug 15, 2019·Scientific Data·Xinyue KangHeng Zheng
Feb 28, 2020·Clinical Microbiology Reviews·Karen Bush, Patricia A Bradford
Jun 22, 2019·The Journal of Antimicrobial Chemotherapy·José-Manuel Ortiz de la RosaLaurent Poirel
Feb 13, 2019·The Journal of Antimicrobial Chemotherapy·Peera Hemarajata, Romney M Humphries
Jan 28, 2018·Expert Opinion on Drug Metabolism & Toxicology·Marco FalconeManjunath Pai
Nov 28, 2018·Antimicrobial Resistance and Infection Control·Wenxia ZhangFupin Hu
Apr 13, 2019·Antibiotics·Federico PerezRobert A Bonomo
Dec 19, 2019·The Journal of Antimicrobial Chemotherapy·Saoussen OueslatiThierry Naas
Dec 16, 2018·Biomolecular NMR Assignments·Jamie VanPeltRichard C Page
Feb 15, 2021·International Journal of Biological Macromolecules·Sourya BhattacharyaSaugata Hazra
Dec 10, 2020·International Journal of Molecular Sciences·Juan C Vázquez-UchaAlejandro Beceiro
Mar 26, 2021·Frontiers in Microbiology·Yanqin HuangZackery P Bulman
Jul 7, 2021·Antimicrobial Agents and Chemotherapy·Jacqueline FindlayPatrice Nordmann
Aug 23, 2021·Diagnostic Microbiology and Infectious Disease·Mustafa SadekPatrice Nordmann
Sep 11, 2021·Folia Microbiologica·Natália Kehl Moreira, Juliana Caierão
Sep 28, 2021·The Journal of Antimicrobial Chemotherapy·Shazad MushtaqDavid M Livermore
Oct 5, 2021·Antimicrobial Agents and Chemotherapy·Agnès B JoussetThierry Naas
Jan 29, 2022·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·Laurent PoirelPatrice Nordmann

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Methods Mentioned

BETA
acylation
MDS
deacylation
PCR
electrophoresis
protein assay

Software Mentioned

ProtParam
ProData SX
Fragment Builder
CDOCKER
ExPASy
Discovery Studio

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