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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Importance of the beta 5-beta 6 Loop for the Structure, Catalytic Efficiency, and Stability of Carbapenem-Hydrolyzing Class D beta-Lactamase Subfamily OXA-143

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Author(s):
Antunes, Victor U. [1] ; Llontop, Edgar E. [2] ; Vasconcelos, Fernanda N. da Costa [3] ; de los Santos, Yossef Lopez [4] ; Oliveira, Ronaldo J. [5] ; Lincopan, Nilton [6] ; Farah, Chuck S. [2] ; Doucet, Nicolas [7, 4] ; Mittermaier, Anthony [8, 7] ; Favaro, Denize C. [1, 8]
Total Authors: 10
Affiliation:
[1] Univ Estadual Campinas, Dept Organ Chem, BR-13083970 Sao Paulo, SP - Brazil
[2] Univ Sao Paulo, Inst Chem, Dept Biochem, Av Prof Lineu Prestes 748, BR-05508000 Sao Paulo, SP - Brazil
[3] Univ Florida, Citrus Res & Educ Ctr, Lake Alfred, FL 33850 - USA
[4] Univ Quebec, INRS, Ctr Armand Frappier Sante Biotechnol, Laval, PQ H7V 1B7 - Canada
[5] Univ Fed Triangulo Mineiro, Inst Ciancias Exatas Nat & Educ, Dept Fis, Lab Biofis Teor, BR-38064200 Uberaba, MG - Brazil
[6] Univ Sao Paulo, Inst Biomed Sci, Dept Microbiol, BR-05508900 Sao Paulo, SP - Brazil
[7] Univ Laval, PROTEO, Quebec City, PQ G1V 0A6 - Canada
[8] McGill Univ, Dept Chem, Montreal, PQ H3A 0G4 - Canada
Total Affiliations: 8
Document type: Journal article
Source: BIOCHEMISTRY; v. 58, n. 34, p. 3604-3616, AUG 27 2019.
Web of Science Citations: 0
Abstract

The class D beta-lactamase OXA-143 has been described as an efficient penicillinase, oxacillinase, and carbapenemase. The D224A variant, known as OXA-231, was described in 2012 as exhibiting less activity toward imipenem and increased oxacillinase activity. Additionally, the P227S mutation was reported as a case of convergent evolution for homologous enzymes. To investigate the impact of both mutations (D224A and P227S), we describe in this paper a deep investigation of the enzymatic activities of these three homologues. OXA-143(P227S) presented enhanced catalytic activity against ampicillin, oxacillins, aztreonam, and carbapenems. In addition, OXA-143(P227S) was the only member capable of hydrolyzing ceftazidime. These enhanced activities were due to a combination of a higher affinity (lower K-m) and a higher turnover number (higher k(cat)). We also determined the crystal structure of apo OXA-231. As expected, the structure of this variant is very similar to the published OXA-143 structure, except for the two M223 conformations and the absence of electron density for three solvent-exposed loop segments. Molecular dynamics calculations showed that both mutants experience higher flexibility compared to that of the wild-type form. Therefore, our results illustrate that D224A and P227S act as deleterious and positive mutations, respectively, within the evolutionary path of the OXA-143 subfamily toward a more efficient carbapenemase. (AU)

FAPESP's process: 17/17303-7 - Structure and function of bacterial secretion systems
Grantee:Shaker Chuck Farah
Support type: Research Projects - Thematic Grants
FAPESP's process: 17/22822-3 - Structural and dynamical origins of catalytic differences between OXA-143 and OXA-231
Grantee:Denize Cristina Favaro
Support type: Scholarships abroad - Research