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CE-ICP-SFMS for the detection of S and Zn in Aeromonas hydrophila Zn-beta-lactamase

(2003) LC GC EUROPE. 16(9). p.616-620
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Abstract
Aeromonads are microorganisms that can cause both human and animal infections and is the known source of hospital-acquired infection. Some Aeromonas strains produce metallo-beta-lactamase enzymes, which are at the origin of beta-lactam resistance in members of this genus. The metallo-beta-lactamases are clinically relevant because of their ability to hydrolyse carbapenem antibiotics, and they also represent a relevant investigational model for studying molecular class B beta-lactamases because of their unique enzymological behaviour.(1,2) The Aeromonas hydrophila metallo-beta-lactamase contains Zn as enzymatic cofactor.(3-5) In this work, Zn bound to the metallo-beta-lactamase is separated from free Zn ions by capillary electrophoresis (CE) and the elements Zn and S are detected and quantified simultaneously by use of an inductively coupled plasma-sector field mass spectrometer (ICP-SFMS) operated at medium mass resolution. The goal of this investigation is to develop a method that allows a fast and accurate determination of the Zn/protein ratio. This ratio can be calculated from the Zn/S ratio. The quantification method is limited to proteins where the sulphur stoichiometry is known. The A. hydrophila Zn-beta-lactamase investigated in this work is known to contain 4 methionines and 1 cysteine. For proteins of which the primary structure is unknown, additional structural information is necessary. In this work A. hydrophila Zn-beta-lactamase is being used as proof of concept. Optimum conditions for CE, ICP-SFMS and their hyphenation were investigated. The use of the sodium salt of phytic acid as buffer additive is discussed into more detail.
Keywords
CAPILLARY-ELECTROPHORESIS, PLASMA-MASS SPECTROMETRY, PHYTIC ACID, METALLOTHIONEIN ISOFORMS, SPECIATION, PROTEINS, RESISTANCE, SEPARATION, RESOLUTION

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Chicago
Van Lierde, Veerle, Karel Strijckmans, M Galleni, Bart Devreese, Jozef Van Beeumen, Frank Vanhaecke, and Luc Moens. 2003. “CE-ICP-SFMS for the Detection of S and Zn in Aeromonas Hydrophila Zn-beta-lactamase.” Lc Gc Europe 16 (9): 616–620.
APA
Van Lierde, V., Strijckmans, K., Galleni, M., Devreese, B., Van Beeumen, J., Vanhaecke, F., & Moens, L. (2003). CE-ICP-SFMS for the detection of S and Zn in Aeromonas hydrophila Zn-beta-lactamase. LC GC EUROPE, 16(9), 616–620.
Vancouver
1.
Van Lierde V, Strijckmans K, Galleni M, Devreese B, Van Beeumen J, Vanhaecke F, et al. CE-ICP-SFMS for the detection of S and Zn in Aeromonas hydrophila Zn-beta-lactamase. LC GC EUROPE. 2003;16(9):616–20.
MLA
Van Lierde, Veerle, Karel Strijckmans, M Galleni, et al. “CE-ICP-SFMS for the Detection of S and Zn in Aeromonas Hydrophila Zn-beta-lactamase.” LC GC EUROPE 16.9 (2003): 616–620. Print.
@article{209924,
  abstract     = {Aeromonads are microorganisms that can cause both human and animal infections and is the known source of hospital-acquired infection. Some Aeromonas strains produce metallo-beta-lactamase enzymes, which are at the origin of beta-lactam resistance in members of this genus. The metallo-beta-lactamases are clinically relevant because of their ability to hydrolyse carbapenem antibiotics, and they also represent a relevant investigational model for studying molecular class B beta-lactamases because of their unique enzymological behaviour.(1,2) The Aeromonas hydrophila metallo-beta-lactamase contains Zn as enzymatic cofactor.(3-5) In this work, Zn bound to the metallo-beta-lactamase is separated from free Zn ions by capillary electrophoresis (CE) and the elements Zn and S are detected and quantified simultaneously by use of an inductively coupled plasma-sector field mass spectrometer (ICP-SFMS) operated at medium mass resolution. The goal of this investigation is to develop a method that allows a fast and accurate determination of the Zn/protein ratio. This ratio can be calculated from the Zn/S ratio. The quantification method is limited to proteins where the sulphur stoichiometry is known. The A. hydrophila Zn-beta-lactamase investigated in this work is known to contain 4 methionines and 1 cysteine. For proteins of which the primary structure is unknown, additional structural information is necessary. In this work A. hydrophila Zn-beta-lactamase is being used as proof of concept. Optimum conditions for CE, ICP-SFMS and their hyphenation were investigated. The use of the sodium salt of phytic acid as buffer additive is discussed into more detail.},
  author       = {Van Lierde, Veerle and Strijckmans, Karel and Galleni, M and Devreese, Bart and Van Beeumen, Jozef and Vanhaecke, Frank and Moens, Luc},
  issn         = {1471-6577},
  journal      = {LC GC EUROPE},
  keyword      = {CAPILLARY-ELECTROPHORESIS,PLASMA-MASS SPECTROMETRY,PHYTIC ACID,METALLOTHIONEIN ISOFORMS,SPECIATION,PROTEINS,RESISTANCE,SEPARATION,RESOLUTION},
  language     = {eng},
  number       = {9},
  pages        = {616--620},
  title        = {CE-ICP-SFMS for the detection of S and Zn in Aeromonas hydrophila Zn-beta-lactamase},
  volume       = {16},
  year         = {2003},
}

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