Proteomic Analysis

  • Protocol
  • First Online:
Pseudomonas Methods and Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1149))

  • 11k Accesses

Abstract

Proteome provides highly valuable information on the amount, modifications, and subcellular localization of polypeptides. Accordingly, geneticists, molecular biologists, and biochemists have logically applied these new tools to respond to different lines of biological questions (inventory of proteins, impact of a mutation, dynamics of protein regulation under a given exposure, …). However, even if the results obtained are very informative, this approach needs an excellent experimental design which ensures robustness and thus yields reproducibility. The present chapter gives appropriate methods for assessing the proteome of Pseudomonas aeruginosa by using a two-dimensional gel electrophoresis approach. Protocols for crude protein extraction, protein separation by using immobilized pH gradients, and protein identification by Liquid Chromatography coupled with tandem Mass Spectrometry (LC-MS/MS) are given.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info
Hardcover Book
USD 299.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Fleischmann RD, Adams MD, White O, Clayton RA, Kirkness EF, Kerlavage AR et al (1995) Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science 269:496–521

    Article  CAS  Google Scholar 

  2. Wilkins MR, Sanchez JC, Gooley AA, Appel RD, Humphrey-Smith I, Hochstrasser DF et al (1995) Progress in proteome projects: why all proteins expressed by a genome should be identified and how to do it. Biotechnol Genet Eng Rev 13:19–50

    Article  Google Scholar 

  3. Arevalo-Ferro C, Buschmann J, Reil G, Gorg A, Wiehlmann L, Tummler B et al (2004) Proteome analysis of intraclonal diversity of two Pseudomonas aeruginosa TB clone isolates. Proteomics 4:1241–1246

    Article  CAS  Google Scholar 

  4. Choi DS, Kim DK, Choi SJ, Lee J, Choi JP, Rho S et al (2011) Proteomic analysis of outer membrane vesicles derived from Pseudomonas aeruginosa. Proteomics 11:3424–3429

    Article  CAS  Google Scholar 

  5. Hare NJ, Solis N, Harmer G, Marzook NB, Rose B, Harbour C et al (2012) Proteomic profiling of Pseudomonas aeruginosa AES-1R, PAO1 and PA14 reveals potential virulence determinants associated with a transmissible cystic fibrosis-associated strain. BMC Microbiol 12:16. doi:10.1186/1471-2180-12-16

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Imperi F, Ciccosanti F, Perdomo AB, Tiburzi F, Mancone C, Alonzi T et al (2009) Analysis of the periplasmic proteome of Pseudomonas aeruginosa, a metabolically versatile opportunistic pathogen. Proteomics 9:1901–1915

    Article  CAS  Google Scholar 

  7. Nouwens AS, Cordwell SJ, Larsen MR, Molloy MP, Gillings M, Willcox MD et al (2000) Complementing genomics with proteomics: the membrane subproteome of Pseudomonas aeruginosa PAO1. Electrophoresis 21:3797–3809

    Article  CAS  Google Scholar 

  8. Vilain S, Cosette P, Hubert M, Lange C, Junter GA, Jouenne T (2004) Comparative proteomic analysis of planktonic and immobilized Pseudomonas aeruginosa cells: a multivariate statistical approach. Anal Biochem 329:120–130

    Article  CAS  Google Scholar 

  9. Patton WF (2002) Detection technologies in proteome analysis. J Chromatogr B771:3–31

    Google Scholar 

  10. Vilain S, Cosette P, Charlionet R, Hubert M, Lange C, Junter GA et al (2001) Substituting Coomassie brilliant blue for bromophenol blue in two dimensional electrophoresis buffers improves the resolution of 2-DE patterns. Electrophoresis 22:4368–4374

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Thierry Jouenne .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Cosette, P., Jouenne, T. (2014). Proteomic Analysis. In: Filloux, A., Ramos, JL. (eds) Pseudomonas Methods and Protocols. Methods in Molecular Biology, vol 1149. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-0473-0_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-0473-0_17

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-0472-3

  • Online ISBN: 978-1-4939-0473-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Navigation