β-lactam antibiotic binding sites in Streptococcus pneumoniae StkP kinase extracellular region / 中华微生物学和免疫学杂志
Chinese Journal of Microbiology and Immunology
;
(12): 556-561, 2022.
Artigo
em Chinês
| WPRIM
| ID: wpr-958225
ABSTRACT
Objective:
To analyze the binding ability of motifs in the serine/threonine kinase StkP extracellular region (EC-StkP) of Streptococcus pneumoniae to β-lactam antibiotics.Methods:
Three motifs (SXXK) in the EC-StkP were mutated into AXXA, respectively or simultaneously. Four mutant plasmids (EC- stkp-AXXA1, EC- stkp-AXXA2, EC- stkp-AXXA3 and EC- stkp-AXXA4) were transfected into recipient cells for cloning and expression. SDS-PAGE combined with gel image analysis was used to detect the expression of the recombinant mutant proteins (EC-rStkP-AXXA1, EC-rStkP-AXXA2, EC-rStkP-AXXA3 and EC-rStkP-AXXA4). The expressed mutated proteins were extracted and purified by Ni-NTA affinity chromatography. The binding abilities of the mutant proteins to penicillin (PCN) and cefotaxime (CTX) were detected by isothermal titration calorimetry (ITC 200) and surface plasmon resonance (Biacore t200).Results:
PCN and CTX could not bind to the expressed proteins with mutations in the first or the third motif (EC-rStkP-AXXA1, EC-rStkP-AXXA3, EC-rStkP-AXXA4). EC-rStkP-AXXA2 could weakly bind to CTX, but not to PCN.Conclusions:
All three motifs in the EC-StkP of Streptococcus pneumoniae could bind to β-lactam antibiotics with the first and the third motifs being more important.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Chinese Journal of Microbiology and Immunology
Ano de publicação:
2022
Tipo de documento:
Artigo
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