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Analysis of the membrane proteome of ciprofloxacin-resistant macrophages by stable isotope labeling with amino acids in cell culture (SILAC)

Nancy E Caceres, Maarten Aerts, Béatrice Marquez, Marie-Paule Mingeot-Leclercq, Paul M Tulkens, Bart Devreese UGent and Françoise Van Bambeke (2013) PLOS ONE. 8(3).
abstract
Overexpression of multidrug transporters is a well-established mechanism of resistance to chemotherapy, but other changes may be co-selected upon exposure to drugs that contribute to resistance. Using a model of J774 macrophages made resistant to the fluoroquinolone antibiotic ciprofloxacin and comparing it with the wild-type parent cell line, we performed a quantitative proteomic analysis using the stable isotope labeling with amino acids in cell culture technology coupled with liquid chromatography electrospray ionization Fourier transform tandem mass spectrometry (LC-ESI-FT-MS/MS) on 2 samples enriched in membrane proteins (fractions F1 and F2 collected from discontinuous sucrose gradient). Nine hundred proteins were identified with at least 3 unique peptides in these 2 pooled fractions among which 61 (F1) and 69 (F2) showed a significantly modified abundance among the 2 cell lines. The multidrug resistance associated protein Abcc4, known as the ciprofloxacin efflux transporter in these cells, was the most upregulated, together with Dnajc3, a protein encoded by a gene located downstream of Abcc4. The other modulated proteins are involved in transport functions, cell adhesion and cytoskeleton organization, immune response, signal transduction, and metabolism. This indicates that the antibiotic ciprofloxacin is able to trigger a pleiotropic adaptative response in macrophages that includes the overexpression of its efflux transporter.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
liquid chromatography electrospray ionization fourier transform tandem mass spectrometry, Dnajc3, macrophages, Fluoroquinolone, Abcc4, membrane proteome, western-blot, NF-KAPPA-B, PHOSPHOLIPID-BINDING PROTEINS, GROWTH-FACTOR RECEPTOR, PROSTATE-CANCER, ENDOPLASMIC-RETICULUM, SUBCELLULAR-LOCALIZATION, MULTIDRUG-RESISTANT, EFFLUX TRANSPORTER, PREVENTS APOPTOSIS, J774 MACROPHAGES
journal title
PLOS ONE
PLoS One
volume
8
issue
3
article number
e58285
pages
15 pages
Web of Science type
Article
Web of Science id
000318334500063
JCR category
MULTIDISCIPLINARY SCIENCES
JCR impact factor
3.534 (2013)
JCR rank
8/55 (2013)
JCR quartile
1 (2013)
ISSN
1932-6203
DOI
10.1371/journal.pone.0058285
language
English
UGent publication?
yes
classification
A1
additional info
the first two authors (NEC and MA) contributed equally to the work
copyright statement
I have retained and own the full copyright for this publication
id
3239987
handle
http://hdl.handle.net/1854/LU-3239987
date created
2013-06-11 20:13:26
date last changed
2016-12-21 15:42:31
@article{3239987,
  abstract     = {Overexpression of multidrug transporters is a well-established mechanism of resistance to chemotherapy, but other changes may be co-selected upon exposure to drugs that contribute to resistance. Using a model of J774 macrophages made resistant to the fluoroquinolone antibiotic ciprofloxacin and comparing it with the wild-type parent cell line, we performed a quantitative proteomic analysis using the stable isotope labeling with amino acids in cell culture technology coupled with liquid chromatography electrospray ionization Fourier transform tandem mass spectrometry (LC-ESI-FT-MS/MS) on 2 samples enriched in membrane proteins (fractions F1 and F2 collected from discontinuous sucrose gradient). Nine hundred proteins were identified with at least 3 unique peptides in these 2 pooled fractions among which 61 (F1) and 69 (F2) showed a significantly modified abundance among the 2 cell lines. The multidrug resistance associated protein Abcc4, known as the ciprofloxacin efflux transporter in these cells, was the most upregulated, together with Dnajc3, a protein encoded by a gene located downstream of Abcc4. The other modulated proteins are involved in transport functions, cell adhesion and cytoskeleton organization, immune response, signal transduction, and metabolism. This indicates that the antibiotic ciprofloxacin is able to trigger a pleiotropic adaptative response in macrophages that includes the overexpression of its efflux transporter.},
  articleno    = {e58285},
  author       = {Caceres, Nancy E and Aerts, Maarten and Marquez, B{\'e}atrice and Mingeot-Leclercq, Marie-Paule and Tulkens, Paul M and Devreese, Bart and Van Bambeke, Fran\c{c}oise},
  issn         = {1932-6203},
  journal      = {PLOS ONE},
  keyword      = {liquid chromatography electrospray ionization fourier transform tandem mass spectrometry,Dnajc3,macrophages,Fluoroquinolone,Abcc4,membrane proteome,western-blot,NF-KAPPA-B,PHOSPHOLIPID-BINDING PROTEINS,GROWTH-FACTOR RECEPTOR,PROSTATE-CANCER,ENDOPLASMIC-RETICULUM,SUBCELLULAR-LOCALIZATION,MULTIDRUG-RESISTANT,EFFLUX TRANSPORTER,PREVENTS APOPTOSIS,J774 MACROPHAGES},
  language     = {eng},
  number       = {3},
  pages        = {15},
  title        = {Analysis of the membrane proteome of ciprofloxacin-resistant macrophages by stable isotope labeling with amino acids in cell culture (SILAC)},
  url          = {http://dx.doi.org/10.1371/journal.pone.0058285},
  volume       = {8},
  year         = {2013},
}

Chicago
Caceres, Nancy E, Maarten Aerts, Béatrice Marquez, Marie-Paule Mingeot-Leclercq, Paul M Tulkens, Bart Devreese, and Françoise Van Bambeke. 2013. “Analysis of the Membrane Proteome of Ciprofloxacin-resistant Macrophages by Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC).” Plos One 8 (3).
APA
Caceres, N. E., Aerts, M., Marquez, B., Mingeot-Leclercq, M.-P., Tulkens, P. M., Devreese, B., & Van Bambeke, F. (2013). Analysis of the membrane proteome of ciprofloxacin-resistant macrophages by stable isotope labeling with amino acids in cell culture (SILAC). PLOS ONE, 8(3).
Vancouver
1.
Caceres NE, Aerts M, Marquez B, Mingeot-Leclercq M-P, Tulkens PM, Devreese B, et al. Analysis of the membrane proteome of ciprofloxacin-resistant macrophages by stable isotope labeling with amino acids in cell culture (SILAC). PLOS ONE. 2013;8(3).
MLA
Caceres, Nancy E, Maarten Aerts, Béatrice Marquez, et al. “Analysis of the Membrane Proteome of Ciprofloxacin-resistant Macrophages by Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC).” PLOS ONE 8.3 (2013): n. pag. Print.