Silac mouse for quantitative proteomics uncovers kindlin-3 as an essential factor for red blood cell function

Krüger, M., Moser, M., Ussar, S., Thievessen, I., Luber, C.A., Forner, F., Schmidt, S., Zanivan, S. , Fässler, R. and Mann, M. (2008) Silac mouse for quantitative proteomics uncovers kindlin-3 as an essential factor for red blood cell function. Cell, 134(2), pp. 353-364. (doi: 10.1016/j.cell.2008.05.033)

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.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Zanivan, Professor Sara
• Authors: Krüger, M., Moser, M., Ussar, S., Thievessen, I., Luber, C.A., Forner, F., Schmidt, S., Zanivan, S., Fässler, R., and Mann, M.
• College/School: College of Medical Veterinary and Life Sciences > School of Cancer Sciences
• Journal Name: Cell
• Publisher: Cell Press
• ISSN: 0092-8674
• ISSN (Online): 1097-4172
• Copyright Holders: Copyright © 2008 Elsevier Inc.
• First Published: First published in Cell 134(2):353-364
• Publisher Policy: Reproduced in accordance with the copyright policy of the publisher