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Applied and Environmental Microbiology, September 2007, p. 5653-5656, Vol. 73, No. 17
0099-2240/07/$08.00+0     doi:10.1128/AEM.00667-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Labeling of Bifidobacterium longum Cells with ¹³C-Substituted Leucine for Quantitative Proteomic Analyses ^,

Yohann Couté,¹ Céline Hernandez,² Ron D. Appel,² Jean-Charles Sanchez,¹ and Abelardo Margolles^1,3*

Biomedical Proteomics Research Group, Department of Structural Biology and Bioinformatics, University of Geneva, 1 Rue Michel Servet, 1211 Geneva 14, Switzerland,¹ Proteome Informatics Group, Swiss Institute of Bioinformatics, Geneva, Switzerland,² Instituto de Productos Lácteos de Asturias, Consejo Superior de Investigaciones Científicas (CSIC), Ctra. Infiesto s/n, 33300 Villaviciosa, Asturias, Spain³

Received 23 March 2007/ Accepted 25 June 2007

Stable isotope labeling of amino acids in cell culture was used for Bifidobacterium longum. A comprehensive proteomic strategy was developed and validated by designing an appropriate semidefined medium that allows stable replacement of natural leucine by [¹³C₆]leucine. Using this strategy, proteins having variations of at least 50% in their expression rates can be quantified with great confidence.

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* Corresponding author. Mailing address: Instituto de Productos Lácteos de Asturias, Consejo Superior de Investigaciones Científicas (CSIC), Ctra. Infiesto s/n, 33300 Villaviciosa, Asturias, Spain. Phone: 34 985 89 21 31. Fax: 34 985 89 22 33. E-mail: amargolles{at}ipla.csic.es

Published ahead of print on 29 June 2007.

Supplemental material for this article may be found at http://aem.asm.org/.

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Applied and Environmental Microbiology, September 2007, p. 5653-5656, Vol. 73, No. 17
0099-2240/07/$08.00+0     doi:10.1128/AEM.00667-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

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