Ghent University Academic Bibliography

Advanced

Mutational analysis of the zinc- and substrate-binding sites in the CphA metallo-beta-lactamase from Aeromonas hydrophila

C Bebrone, C Anne, F Kerff, G Garau, Kris De Vriendt, R Lantin, Bart Devreese UGent, Jozef Van Beeumen, O Dideberg, JM Frere, et al. (2008) Biochemical Journal. 414(1). p.151-159
abstract
The subclass B2 CphA (Carbapenemase hydrolysing Aeromonas) beta-lactamase from Aeromonas hydrophila is a Zn(2+)-containing enzyme that specifically hydrolyses carbapenems. In an effort to evaluate residues potentially involved in metal binding and/or catalysis (His(118), Asp(120), His(196) and His(263)) and in substrate specificity (Val(67), Thr(157), Lys(224) and Lys(226)), site-directed mutants of CphA were generated and characterized. Our results confirm that the first zinc ion is in interaction with Asp(120) and His(263), and thus is located in the 'cysteine' zinc-binding site. His(118) and His(196) residues seem to be interacting with the second zinc ion, as their replacement by alanine residues has a negative effect on the affinity for this second metal ion. Val(67) plays a significant role in the binding of biapenem and benzylpenicillin. The properties of a mutant with a five residue (LFKHV) insertion just after Val(67) also reveals the importance of this region for substrate binding. This latter mutant has a higher affinity for the second zinc ion than wild-type CphA. The T157A mutant exhibits a significantly modified activity spectrum. Analysis of the K224Q and N116H/N220G/K224Q mutants suggests a significant role for Lys(224) in the binding of substrate. Lys(226) is not essential for the binding and hydrolysis of substrates. Thus the present paper helps to elucidate the position of the second zinc ion, which was controversial, and to identify residues important for substrate binding.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
journal title
Biochemical Journal
Biochem. J.
volume
414
issue
1
pages
151 - 159
Web of Science type
Article
Web of Science id
000258542100015
JCR category
BIOCHEMISTRY & MOLECULAR BIOLOGY
JCR impact factor
4.371 (2008)
JCR rank
72/276 (2008)
JCR quartile
1 (2008)
ISSN
0264-6021
DOI
10.1042/BJ20080375
language
English
UGent publication?
yes
classification
A1
id
531651
handle
http://hdl.handle.net/1854/LU-531651
date created
2009-03-26 13:31:07
date last changed
2016-12-19 15:45:42
@article{531651,
  abstract     = {The subclass B2 CphA (Carbapenemase hydrolysing Aeromonas) beta-lactamase from Aeromonas hydrophila is a Zn(2+)-containing enzyme that specifically hydrolyses carbapenems. In an effort to evaluate residues potentially involved in metal binding and/or catalysis (His(118), Asp(120), His(196) and His(263)) and in substrate specificity (Val(67), Thr(157), Lys(224) and Lys(226)), site-directed mutants of CphA were generated and characterized. Our results confirm that the first zinc ion is in interaction with Asp(120) and His(263), and thus is located in the 'cysteine' zinc-binding site. His(118) and His(196) residues seem to be interacting with the second zinc ion, as their replacement by alanine residues has a negative effect on the affinity for this second metal ion. Val(67) plays a significant role in the binding of biapenem and benzylpenicillin. The properties of a mutant with a five residue (LFKHV) insertion just after Val(67) also reveals the importance of this region for substrate binding. This latter mutant has a higher affinity for the second zinc ion than wild-type CphA. The T157A mutant exhibits a significantly modified activity spectrum. Analysis of the K224Q and N116H/N220G/K224Q mutants suggests a significant role for Lys(224) in the binding of substrate. Lys(226) is not essential for the binding and hydrolysis of substrates. Thus the present paper helps to elucidate the position of the second zinc ion, which was controversial, and to identify residues important for substrate binding.},
  author       = {Bebrone, C and Anne, C and Kerff, F and Garau, G and De Vriendt, Kris and Lantin, R and Devreese, Bart and Van Beeumen, Jozef and Dideberg, O and Frere, JM and Galleni, M},
  issn         = {0264-6021},
  journal      = {Biochemical Journal},
  language     = {eng},
  number       = {1},
  pages        = {151--159},
  title        = {Mutational analysis of the zinc- and substrate-binding sites in the CphA metallo-beta-lactamase from Aeromonas hydrophila},
  url          = {http://dx.doi.org/10.1042/BJ20080375},
  volume       = {414},
  year         = {2008},
}

Chicago
Bebrone, C, C Anne, F Kerff, G Garau, Kris De Vriendt, R Lantin, Bart Devreese, et al. 2008. “Mutational Analysis of the Zinc- and Substrate-binding Sites in the CphA Metallo-beta-lactamase from Aeromonas Hydrophila.” Biochemical Journal 414 (1): 151–159.
APA
Bebrone, C., Anne, C., Kerff, F., Garau, G., De Vriendt, K., Lantin, R., Devreese, B., et al. (2008). Mutational analysis of the zinc- and substrate-binding sites in the CphA metallo-beta-lactamase from Aeromonas hydrophila. Biochemical Journal, 414(1), 151–159.
Vancouver
1.
Bebrone C, Anne C, Kerff F, Garau G, De Vriendt K, Lantin R, et al. Mutational analysis of the zinc- and substrate-binding sites in the CphA metallo-beta-lactamase from Aeromonas hydrophila. Biochemical Journal. 2008;414(1):151–9.
MLA
Bebrone, C, C Anne, F Kerff, et al. “Mutational Analysis of the Zinc- and Substrate-binding Sites in the CphA Metallo-beta-lactamase from Aeromonas Hydrophila.” Biochemical Journal 414.1 (2008): 151–159. Print.