In vivo quantitative proteomics: the SILAC mouse.

Area of Science:

• Proteomics
• Biochemistry
• Animal Models

Background:

• Mass spectrometry-based proteomics offers in-depth proteome characterization.
• Stable isotope labeling with amino acid in cell culture (SILAC) is established for quantitative cell line studies.
• Extending SILAC to in vivo models requires robust methodologies.

Purpose of the Study:

• To adapt and validate Stable isotope labeling with amino acid in cell culture (SILAC) for in vivo quantitative proteomic analysis in mice.
• To establish a practical method for generating and maintaining a SILAC mouse colony.
• To optimize strategies for in vivo quantitative proteomic experiments using SILAC mice.

Main Methods:

• Generation and maintenance of a colony of C57BL/6 mice fully labeled with (13)C(6)-Lysine (Lys6).
• Application of SILAC mice technology to quantitatively map proteomic changes in genetically modified mouse models.
• Mass spectrometry-based proteomics for high-throughput protein identification and quantification.

Main Results:

• Successful generation and maintenance of a SILAC mouse colony.
• Quantitative proteomic analysis confirmed the absence of targeted proteins (β1 integrin, β-Parvin, Kindlin-3).
• Identification of a novel role for Kindlin-3 in red blood cells.

Conclusions:

• SILAC mice technology is a powerful tool for in vivo quantitative proteomics.
• This methodology enables the study of protein function and interactions in complex animal systems.
• The developed methods provide a practical framework for future in vivo proteomic research.