Protein metabolic labeling

Original review on protein metabolic labelling for proteomics study.

By Robert J. Beynon{ddagger} and Julie M. Pratt, Molecular & Cellular Proteomics 4:857-872, 2005

There is a gradual emergence of the realisation of the advantages of stable isotope labelling for proteomics. However, many stable isotope label approaches rely on labelling in vitro using complex and sometimes expensive reagents. This review discusses strategies for labelling protein in vivo, through metabolic incorporation of label into protein. This approach has many advantages, is particularly suited to single cells grown in culture (prokaryotic or eukaryotic) but is nonetheless subject to a number of complicating factors that must be controlled in order that meaningful experiments can be conducted. Confounding issues include the metabolic lability of the amino acid precursor, incomplete labelling, and the role of protein turnover in labelling kinetics. All of these are controllable, provided that appropriate precautions are dopted.

Keywords: stable isotopes, amino acid precursors, SILAC, protein turnover, proteome dynamics, mass spectrometry, post-translational modifications

Recent technique papers on metabolic labeling of proteins.

1: Engelsberger WR, Erban A, Kopka J, Schulze WX.
Metabolic labeling of plant cell cultures with K15NO3 as a tool for quantitative
analysis of proteins and metabolites.
Plant Methods. 2006 Sep 4;2:14.

2: Huttlin EL, Hegeman AD, Harms AC, Sussman MR.
Comparison of full versus partial metabolic labeling for quantitative proteomics
analysis in Arabidopsis thaliana.
Mol Cell Proteomics. 2007 May;6(5):860-81. Epub 2007 Feb 9.

3: Beynon RJ, Pratt JM.
Metabolic labeling of proteins for proteomics.
Mol Cell Proteomics. 2005 Jul;4(7):857-72. Epub 2005 Apr 22. Review.

4: Snijders AP, de Vos MG, Wright PC.
Novel approach for peptide quantitation and sequencing based on 15N and 13C
metabolic labeling.
J Proteome Res. 2005 Mar-Apr;4(2):578-85.

5: Millecamps S, Julien JP.
[35S]Methionine metabolic labeling to study axonal transport of neuronal
intermediate filament proteins in vivo.
Methods Cell Biol. 2004;78:555-71. No abstract available.

6: Gehrmann ML, Hathout Y, Fenselau C.
Evaluation of metabolic labeling for comparative proteomics in breast cancer
cells.
J Proteome Res. 2004 Sep-Oct;3(5):1063-8.

7: Hang HC, Yu C, Kato DL, Bertozzi CR.
A metabolic labeling approach toward proteomic analysis of mucin-type O-linked
glycosylation.
Proc Natl Acad Sci U S A. 2003 Dec 9;100(25):14846-51. Epub 2003 Dec 1.

Archived page

Last update 17-Jun-2007, Rating Good of 0 votes.