Biochemical, structural, and computational studies of a γ-carbonic anhydrase from the pathogenic bacterium Burkholderia pseudomallei.
Comput Struct Biotechnol J
; 20: 4185-4194, 2022.
Article
in En
| MEDLINE
| ID: mdl-36016712
ABSTRACT
Melioidosis is a severe disease caused by the highly pathogenic gram-negative bacterium Burkholderia pseudomallei. Several studies have highlighted the broad resistance of this pathogen to many antibiotics and pointed out the pivotal importance of improving the pharmacological arsenal against it. Since γ-carbonic anhydrases (γ-CAs) have been recently introduced as potential and novel antibacterial drug targets, in this paper, we report a detailed characterization of BpsγCA, a γ-CA from B. pseudomallei by a multidisciplinary approach. In particular, the enzyme was recombinantly produced and biochemically characterized. Its catalytic activity at different pH values was measured, the crystal structure was determined and theoretical pKa calculations were carried out. Results provided a snapshot of the enzyme active site and dissected the role of residues involved in the catalytic mechanism and ligand recognition. These findings are an important starting point for developing new anti-melioidosis drugs targeting BpsγCA.
BME, 2-betamercaptoethanol; BSA, Bovine Serum Albumin; BpsßCA, ß-CA from B. pseudomallei; BpsγCA, γ-CA from B. pseudomallei; Burkholderia pseudomallei; CA, Carbonic Anhydrase; CA_D, γ-CA from Discovery Deep Brine Pool; CCD, Charge Coupled Device; CD, circular dichroism; Cag, γ-CA from G. kaustophilus; Cam, γ-CA from M. thermophila; Carbonic anhydrases; Crystal structure; Cyt C, horse Cytochrome C; Melioidosis; PDB, Protein Data Bank; PEG, Polyethylene glycol; PSR, proton shuttle residue; RicA, γ-CA from B. abortus; SEC, size exclusion chromatography; TeCcmM, γ-CA from T. elongatus; TtCA, γ-CA from T. thermophilus HB8; Yrda, γ-CA from E. coli; Zn-Cap, γ-CA from P. horikoshii; pKa calculations; r.s.m.d., root mean square deviation
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
Comput Struct Biotechnol J
Year:
2022
Document type:
Article
Affiliation country:
Italy