The Scientific World Journal

The Scientific World Journal / 2002 / Article

Research Article | Open Access

Volume 2 |Article ID 564140 | https://doi.org/10.1100/tsw.2002.215

Irena Fonda, Maja Kenig, Vladka Gaberc-Porekar, Primo Pristovaek, Viktor Menart, "Attachment of Histidine Tags to Recombinant Tumor Necrosis Factor-Alpha Drastically Changes Its Properties", The Scientific World Journal, vol. 2, Article ID 564140, 14 pages, 2002. https://doi.org/10.1100/tsw.2002.215

Attachment of Histidine Tags to Recombinant Tumor Necrosis Factor-Alpha Drastically Changes Its Properties

Received04 Dec 2001
Revised07 Mar 2002
Accepted14 Mar 2002

Abstract

When studying two different histidine tags attached to the N-termini of the trimeric cytokine tumor necrosis factor alpha (TNF), the biological activity — measured as cytotoxicity on the L-929 cell line — of both tagged proteins was drastically reduced. The longer His10 tag reduced cytotoxicity to approximately 16% and the shorter His7 tag to 6% of the activity of their nontagged counterparts. After removal of the tags, biological activities reverted to the expected normal values, which clearly shows the key role of the attached histidine tags in diminishing biological activity. Studies on the mechanism of these effects revealed no specific interactions and showed that even the natural flexible N-terminus of TNF presents a steric hindrance for receptor binding, while any extension of the N-terminus increases this hindrance and consequently reduces biological activity. Also, in other proteins, the ligand or substrate binding sites may be hindered by histidine tags, leading to wrong conclusions about biological activity or other properties of the proteins. Thus caution is advised when using His-tagged proteins directly in screening procedures or in research.


More related articles

 PDF Download Citation Citation
 Order printed copiesOrder
Views92
Downloads1401
Citations

Related articles

We are committed to sharing findings related to COVID-19 as quickly as possible. We will be providing unlimited waivers of publication charges for accepted research articles as well as case reports and case series related to COVID-19. Review articles are excluded from this waiver policy. Sign up here as a reviewer to help fast-track new submissions.