HA-tag explained

The HA-tag is a protein tag derived from the human influenza hemagglutinin (HA) protein, which allows the virus to target and enter host cells. An HA-tag is composed of a peptide derived from the HA-molecule corresponding to amino acids 98-106, which can be recognized and selectively bound by commercially available antibodies. This makes HA a powerful tool in molecular biology, commonly included in expression vectors and in the production of recombinant proteins.[1] Like other epitope tags, HA-tag is small and generally does not alter the traits of proteins it is attached to.[2] As a result HA-tags are often used to identify protein-protein interactions or to detect protein expression, using Co-Immunoprecipitation or Western blot respectively.[3] [4]

The HA-tag is not suitable for detection or purification of proteins from apoptotic cells since it is cleaved by Caspase-3 and / or Caspase-7 after its sequence DVPD, causing it to lose its immunoreactivity.[5] Labeling of endogenous proteins with HA-tag using CRISPR was recently accomplished in-vivo in differentiated neurons.[6]

Sequence

The DNA sequences for the HA-tag include: 5'-TAC-CCA-TAC-GAT-GTT-CCA-GAT-TAC-GCT-3' or 5'-TAT-CCA-TAT-GAT-GTT-CCA-GAT-TAT-GCT-3'. The resulting amino acid sequence is YPYDVPDYA (Tyr-Pro-Tyr-Asp-Val-Pro-Asp-Tyr-Ala).[7]

See also

Further reading

Notes and References

  1. Field J, Nikawa J, Broek D, MacDonald B, Rodgers L, Wilson IA, Lerner RA, Wigler M . Purification of a RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method. . Mol Cell Biol . 8 . 5 . 2159–65 . 1988 . 2455217 . 363397 . 10.1128/mcb.8.5.2159-2165.1988 .
  2. Kimple . Michelle E. . Brill . Allison L. . Pasker . Renee L. . 2013-09-24 . Overview of Affinity Tags for Protein Purification . Current Protocols in Protein Science . 73 . Unit–9.9 . 10.1002/0471140864.ps0909s73 . 1934-3655 . 4527311 . 24510596.
  3. Web site: Anti-HA Tag Antibody Updated 28/10/2021. Merck Millipore.
  4. Web site: Overview of Epitope Tagging - US . 2024-07-25 . www.thermofisher.com . en.
  5. Schembri L, Dalibart R, Tomasello F, Legembre P, Ichas F, De Giorgi F . The HA tag is cleaved and loses immunoreactivity during apoptosis . Nature Methods . 4 . 2 . 107–8 . February 2007 . 17264856 . 10.1038/nmeth0207-107 . 32173039 .
  6. Mikuni T, Nishiyama J, Sun Y, Kamasawa N, Yasuda R . High Throughput, High Resolution Mapping of Protein Localization in Mammalian Brain by In Vivo Genome Editing . Cell . 2016 . 165 . 7 . 1803–1817 . 27180908 . 10.1016/j.cell.2016.04.044 . 4912470 .
  7. Web site: Addgene: HA-tag . 2024-07-25 . www.addgene.org . en.

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