The tandem affinity purification (TAP) method: a general procedure of protein complex purification

Area of Science:

• Molecular Biology
• Biochemistry
• Proteomics

Background:

• Purifying biological complexes to homogeneity is crucial for component identification.
• Existing purification methods lack a universal strategy, varying significantly between proteins.

Purpose of the Study:

• To develop a generalizable method for rapid purification of protein complexes under native conditions.
• To enable proteome exploration and protein purification without prior knowledge of complex composition or function.

Main Methods:

• The tandem affinity purification (TAP) method was developed, involving fusion of a TAP tag to the target protein.
• Purification is achieved under native conditions, starting from a small number of cells.
• The method allows for variations to purify specific complexes or subtract undesired ones.

Main Results:

• The TAP method enables rapid purification of active macromolecular complexes at their natural expression levels.
• It requires no prior knowledge of complex composition or function.
• The method is adaptable to various organisms, initially developed in yeast.

Conclusions:

• The TAP method offers a simple, high-yield, and widely applicable procedure for protein purification.
• It is a valuable tool for proteome exploration and identifying components of biological complexes.
• Its versatility makes it suitable for diverse research applications in molecular biology.