Crystallization and Preliminary X-Ray Crystallographic Analysis of CTXM- 15, an Extended-spectrum β-Lactamase Conferring Worldwide Emerging Antibiotic Resistance

Young      Jun An; Jung      Hun Lee; Ha      Il Jung; Seung      Ghyu Sohn; Jae      Jin Lee; Kwang      Seung Park; Xing      Wu; Byeong      Chul Jeong; Choong-Min      Kang; Sun-Shin      Cha; Sang      Hee Lee

doi:10.2174/092986611796011400

Abstract

CTX-M-15, an extended-spectrum β-lactamase emerging worldwide, hydrolyzes lactam ring of β-lactam antibiotics, and thus causes therapeutic failure and a lack of eradication of pathogenic bacteria by third-generation β-lactams. Therefore, the enzyme is a potential target for developing agents against pathogens isolated from patients suffering from nosocomial infections. The CTX-M-15 protein was purified and crystallized at 298 K. X-ray diffraction data from CTXM- 15 crystal have been collected to 1.46 Å resolution using synchrotron radiation. The crystal of CTX-M-15 belongs to space group P212121, with unit-cell parameters a = 45.50, b = 44.23, and c = 116.92 Å. Analysis of the packing density shows that the asymmetric unit probably contains two molecules with a solvent content of 41.26%.

Keywords: Cephalosporins, variants, oxyimino β-lactams, ESBLs, antibiotics, polyacrylamide SDS gel, Polyethylene glycol, Polymerase chain reaction, Asp240Gly mutation, liquid nitrogen, Escherichia coli, synchrotron radiation, nosocomial infections, third-generation β-lactams, lactam ring, extended-spectrum β-lactamase, CTX-M-15, crystal, ceftazidime, cefotaxime, Antibiotic resistanceCephalosporins, variants, oxyimino β-lactams, ESBLs, antibiotics, polyacrylamide SDS gel, Polyethylene glycol, Polymerase chain reaction, Asp240Gly mutation, liquid nitrogen, Escherichia coli, synchrotron radiation, nosocomial infections, third-generation β-lactams, lactam ring, extended-spectrum β-lactamase, CTX-M-15, crystal, ceftazidime, cefotaxime, Antibiotic resistance