Protein tag
A protein tag, also known as a fusion tag or an affinity tag, is a short peptide sequence that is genetically fused to a target protein. The addition of a protein tag to a protein of interest aims to facilitate its purification, detection, localization, and/or characterization, making protein tags invaluable tools in Molecular biology and Biotechnology research.
History
The concept of protein tags emerged in the early 1980s, addressing the challenge of isolating and studying specific proteins efficiently. The pioneering protein tag developed was the hexahistidine tag (His-tag), comprising six consecutive histidine residues with a high affinity for metal ions like nickel or cobalt. Subsequently, a diverse array of protein tags have been developed, each offering unique properties and applications.
Purpose
Protein tags are versatile tools serving multiple roles in protein research:
- Purification: Tags like the His-tag, glutathione S-transferase (GST) tag, and streptavidin-binding peptide (SBP) tag enable the efficient purification of tagged proteins from complex mixtures.
- Detection: Detection and quantification are facilitated by tags such as the green fluorescent protein (GFP) tag, FLAG tag, and hemagglutinin (HA) tag, which are recognizable by specific antibodies or reagents.
- Localization: Fluorescent protein tags (e.g., GFP, mCherry) allow researchers to observe the subcellular distribution of proteins in live cells via fluorescence microscopy.
- Characterization: Tags such as the maltose-binding protein (MBP) tag and calmodulin-binding peptide (CBP) tag help explore protein structure, function, and interactions.
Commonly Used Protein Tags
The scientific community frequently utilizes several protein tags due to their effectiveness and adaptability:
- His-tag: Binds to metal ions for purification via immobilized metal affinity chromatography (IMAC).
- GST-tag: Derived from the glutathione S-transferase enzyme, it facilitates affinity purification.
- FLAG-tag: A small peptide sequence used extensively for detection and immunoprecipitation.
- GFP-tag: Allows visualization of proteins in live cells through fluorescence microscopy.
- HA-tag: Derived from influenza virus hemagglutinin, useful for detection and immunoprecipitation.
Categories
Protein tags can be categorized by their properties and applications:
- 1. Affinity Tags: Facilitate purification through specific binding interactions. Examples: His-tag, GST-tag, SBP-tag.
- 2. Fluorescent Tags: Enable visualization via fluorescence microscopy. Examples: GFP-tag, mCherry-tag.
- 3. Epitope Tags: Recognized by specific antibodies for detection and immunoprecipitation. Examples: FLAG-tag, HA-tag.
- 4. Enzyme Tags: Derived from enzymes, these tags have diverse applications, such as in protein-protein interaction studies. Examples: MBP-tag, CBP-tag.
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Contributors: Prab R. Tumpati, MD