SnAvi--a new tandem tag for high-affinity protein-complex purification

Area of Science:

• Proteomics and Systems Biology
• Molecular Biology
• Cell Biology

Background:

• Systematic tandem-affinity-purification (TAP) has been highly successful in yeast for understanding protein function.
• Adapting TAP methods for other model organisms, like Caenorhabditis elegans, presents challenges.
• A cell-type-specific TAP system is needed for high-throughput analysis in C. elegans.

Purpose of the Study:

• To develop a novel in vivo fluorescent tag for cell-type-specific tandem affinity purification (TAP) in Caenorhabditis elegans.
• To establish a versatile tool applicable across different model organisms and experimental techniques.

Main Methods:

• Development of the SnAvi-Tag by combining in vivo biotinylation, streptavidin, and a novel SB1 monoclonal antibody epitope.
• Application of the SnAvi-Tag in Escherichia coli, vertebrate cells, and C. elegans.
• Utilizing the tag for tandem affinity purification, western blotting, and in vivo sub-cellular localization.

Main Results:

• The SnAvi-Tag demonstrated versatile application in multiple model systems.
• Successful tandem affinity purification of protein complexes was achieved using the SnAvi-Tag.
• In vivo sub-cellular localization of labeled proteins was successfully performed.

Conclusions:

• The SnAvi-Tag is a novel and versatile in vivo fluorescent tag for protein complex analysis.
• This method facilitates cell-type-specific proteomics and systems biology studies in C. elegans and other organisms.
• The SnAvi-Tag expands the toolkit for molecular and cell biology research.