Ghent University Academic Bibliography

Advanced

Capillary electrophoresis hyphenated to inductively coupled plasma-sector field-mass spectrometry for the stoichiometric determination of Zn bound to Aeromonas hydrophila Zn beta-lactamase

Veerle Van Lierde, Cyrille C Chéry, Karel Strijckmans UGent, Moreno Galleni, Bart Devreese UGent, Jozef Van Beeumen UGent, Luc Moens UGent and Frank Vanhaecke UGent (2004) JOURNAL OF ANALYTICAL ATOMIC SPECTROMETRY. 19(7). p.888-893
abstract
A method was developed for the determination of the stoichiometric Zn/protein ratio for the Aeromonas hydrophila metallo beta-lactamase AE036. Capillary electrophoresis (CE) was used to separate free Zn ions from the Zn bound to the metalloprotein, the only Zn-species which is biochemically relevant. An inductively coupled plasma-sector field-mass spectrometer (ICP-SF-MS), operated at a mass resolution of 3000, was used as an element-specific detector. If the primary structure of the protein and thus the number of sulfur-containing amino acids is known, as is the case for Aeromonas hydrophila Zn beta-lactamase, monitoring of both Zn and S with ICP-SF-MS permits the Zn/protein ratio to be determined. The method was optimised in order to obtain short migration times (less than 10 min) for the metallo beta-lactamase and for standards with bare fused silica capillaries. After optimisation, a 75 mmol L-1 bis-Tris background electrolyte buffered at a pH of 6.75 was used for the separation, a voltage of 30 kV was applied over a capillary with a total length of approximately 70 cm and the sample was introduced hydrodynamically at 1000 mbar s. Quantitative results (stoichiometric Zn/protein ratio of 1 and 2, before and after saturation of the enzyme with Zn, respectively) were obtained using external calibration with albumin as a S standard and ZnCl2 as a Zn standard. The method was also demonstrated to be precise with < 5% RSD on the Zn/protein ratio for n=3. An absolute limit of detection of 8 ng of A. hydrophila Zn beta-lactamase (calculated on the basis of sulfur) was obtained under these conditions.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
METALLOTHIONEIN ISOFORMS, PHYTIC ACID, SPECIATION, PROTEINS, RESISTANCE, SEPARATION, RESOLUTION
journal title
JOURNAL OF ANALYTICAL ATOMIC SPECTROMETRY
J. Anal. At. Spectrom.
volume
19
issue
7
pages
888 - 893
Web of Science type
Article
Web of Science id
000222472700011
JCR category
CHEMISTRY, ANALYTICAL
JCR impact factor
3.926 (2004)
JCR rank
3/70 (2004)
JCR quartile
1 (2004)
ISSN
0267-9477
DOI
10.1039/b400962b
language
English
UGent publication?
yes
classification
A1
id
293910
handle
http://hdl.handle.net/1854/LU-293910
date created
2004-08-19 17:11:00
date last changed
2012-11-12 15:12:15
@article{293910,
  abstract     = {A method was developed for the determination of the stoichiometric Zn/protein ratio for the Aeromonas hydrophila metallo beta-lactamase AE036. Capillary electrophoresis (CE) was used to separate free Zn ions from the Zn bound to the metalloprotein, the only Zn-species which is biochemically relevant. An inductively coupled plasma-sector field-mass spectrometer (ICP-SF-MS), operated at a mass resolution of 3000, was used as an element-specific detector. If the primary structure of the protein and thus the number of sulfur-containing amino acids is known, as is the case for Aeromonas hydrophila Zn beta-lactamase, monitoring of both Zn and S with ICP-SF-MS permits the Zn/protein ratio to be determined. The method was optimised in order to obtain short migration times (less than 10 min) for the metallo beta-lactamase and for standards with bare fused silica capillaries. After optimisation, a 75 mmol L-1 bis-Tris background electrolyte buffered at a pH of 6.75 was used for the separation, a voltage of 30 kV was applied over a capillary with a total length of approximately 70 cm and the sample was introduced hydrodynamically at 1000 mbar s. Quantitative results (stoichiometric Zn/protein ratio of 1 and 2, before and after saturation of the enzyme with Zn, respectively) were obtained using external calibration with albumin as a S standard and ZnCl2 as a Zn standard. The method was also demonstrated to be precise with {\textlangle} 5\% RSD on the Zn/protein ratio for n=3. An absolute limit of detection of 8 ng of A. hydrophila Zn beta-lactamase (calculated on the basis of sulfur) was obtained under these conditions.},
  author       = {Van Lierde, Veerle and Ch{\'e}ry, Cyrille C and Strijckmans, Karel and Galleni, Moreno and Devreese, Bart and Van Beeumen, Jozef and Moens, Luc and Vanhaecke, Frank},
  issn         = {0267-9477},
  journal      = {JOURNAL OF ANALYTICAL ATOMIC SPECTROMETRY},
  keyword      = {METALLOTHIONEIN ISOFORMS,PHYTIC ACID,SPECIATION,PROTEINS,RESISTANCE,SEPARATION,RESOLUTION},
  language     = {eng},
  number       = {7},
  pages        = {888--893},
  title        = {Capillary electrophoresis hyphenated to inductively coupled plasma-sector field-mass spectrometry for the stoichiometric determination of Zn bound to Aeromonas hydrophila Zn beta-lactamase},
  url          = {http://dx.doi.org/10.1039/b400962b},
  volume       = {19},
  year         = {2004},
}

Chicago
Van Lierde, Veerle, Cyrille C Chéry, Karel Strijckmans, Moreno Galleni, Bart Devreese, Jozef Van Beeumen, Luc Moens, and Frank Vanhaecke. 2004. “Capillary Electrophoresis Hyphenated to Inductively Coupled Plasma-sector Field-mass Spectrometry for the Stoichiometric Determination of Zn Bound to Aeromonas Hydrophila Zn Beta-lactamase.” Journal of Analytical Atomic Spectrometry 19 (7): 888–893.
APA
Van Lierde, V., Chéry, C. C., Strijckmans, K., Galleni, M., Devreese, B., Van Beeumen, J., Moens, L., et al. (2004). Capillary electrophoresis hyphenated to inductively coupled plasma-sector field-mass spectrometry for the stoichiometric determination of Zn bound to Aeromonas hydrophila Zn beta-lactamase. JOURNAL OF ANALYTICAL ATOMIC SPECTROMETRY, 19(7), 888–893.
Vancouver
1.
Van Lierde V, Chéry CC, Strijckmans K, Galleni M, Devreese B, Van Beeumen J, et al. Capillary electrophoresis hyphenated to inductively coupled plasma-sector field-mass spectrometry for the stoichiometric determination of Zn bound to Aeromonas hydrophila Zn beta-lactamase. JOURNAL OF ANALYTICAL ATOMIC SPECTROMETRY. 2004;19(7):888–93.
MLA
Van Lierde, Veerle, Cyrille C Chéry, Karel Strijckmans, et al. “Capillary Electrophoresis Hyphenated to Inductively Coupled Plasma-sector Field-mass Spectrometry for the Stoichiometric Determination of Zn Bound to Aeromonas Hydrophila Zn Beta-lactamase.” JOURNAL OF ANALYTICAL ATOMIC SPECTROMETRY 19.7 (2004): 888–893. Print.