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Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability.

ABSTRACT: For years, the use of polyhistidine tags (His-tags) has been a staple in the isolation of recombinant proteins in immobilized metal affinity chromatography experiments. Their usage has been widely beneficial in increasing protein purity from crude cell lysates. For some recombinant proteins, a consequence of His-tag addition is that it can affect protein function and stability. Functional proteins are essential in the elucidation of their biological, kinetic, structural, and thermodynamic properties. In this study, we determine the effect of N-terminal His-tags on the thermal stability of select proteins using differential scanning fluorimetry and identify that the removal of the His-tag can have both beneficial and deleterious effects on their stability.

SUBMITTER: Booth WT

PROVIDER: S-EPMC5793033 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature

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Impact of an N-terminal Polyhistidine Tag on Protein Thermal Stability.

Booth William T WT Schlachter Caleb R CR Pote Swanandi S Ussin Nikita N Mank Nicholas J NJ Klapper Vincent V Offermann Lesa R LR Tang Chuanbing C Hurlburt Barry K BK Chruszcz Maksymilian M

ACS omega 20180122 1

For years, the use of polyhistidine tags (His-tags) has been a staple in the isolation of recombinant proteins in immobilized metal affinity chromatography experiments. Their usage has been widely beneficial in increasing protein purity from crude cell lysates. For some recombinant proteins, a consequence of His-tag addition is that it can affect protein function and stability. Functional proteins are essential in the elucidation of their biological, kinetic, structural, and thermodynamic proper ...[more]

PMID: 29399652

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