SILAC Mouse for Quantitative Proteomics Uncovers Kindlin-3 as an Essential Factor for Red Blood Cell Function

Marcus Krüger, Markus Moser, Siegfried Ussar, Ingo Thievessen, Christian A. Luber, Francesca Forner, Sarah Schmidt, Sara Zanivan, Reinhard Fässler, Matthias Mann

Research output: Contribution to journalArticlepeer-review

560 Scopus citations

Abstract

Stable isotope labeling by amino acids in cell culture (SILAC) has become a versatile tool for quantitative, mass spectrometry (MS)-based proteomics. Here, we completely label mice with a diet containing either the natural or the 13C6-substituted version of lysine. Mice were labeled over four generations with the heavy diet, and development, growth, and behavior were not affected. MS analysis of incorporation levels allowed for the determination of incorporation rates of proteins from blood cells and organs. The F2 generation was completely labeled in all organs tested. SILAC analysis from various organs lacking expression of β1 integrin, β-Parvin, or the integrin tail-binding protein Kindlin-3 confirmed their absence and disclosed a structural defect of the red blood cell membrane skeleton in Kindlin-3-deficient erythrocytes. The SILAC-mouse approach is a versatile tool by which to quantitatively compare proteomes from knockout mice and thereby determine protein functions under complex in vivo conditions.

Original languageEnglish
Pages (from-to)353-364
Number of pages12
JournalCell
Volume134
Issue number2
DOIs
StatePublished - 25 Jul 2008
Externally publishedYes

Keywords

  • PROTEINS
  • SYSBIO

Fingerprint

Dive into the research topics of 'SILAC Mouse for Quantitative Proteomics Uncovers Kindlin-3 as an Essential Factor for Red Blood Cell Function'. Together they form a unique fingerprint.

Cite this