Protein structure modeling indicates hexahistidine‐tag interference with enzyme activity

Protein structure modeling indicates hexahistidine‐tag interference with enzyme activity Unusual kinetic characteristics of tropinone reductase, an enzyme in the family of short chain dehydrogenases, prompted to investigate a possible impact of the hexahistidine affinity tag on catalytic properties. Comparison of enzymes from Solanum dulcamara, Solanaceae, tagged at the N‐terminus or at the C‐terminus revealed that the C‐terminally tagged form was functionally impaired. Protein modeling indicated that the hexahistidine tag attached at the C‐terminus but not at the N‐terminus of the polypeptide can interfere with the active site by steric or electrostatic interactions. In consequence, protein modeling is suggested before enzyme expression with affinity tags to estimate possible interactions of affinity tags with the active center. Proteins 2008. © 2008 Wiley‐Liss, Inc. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Proteins: Structure Function and Bioinformatics Wiley

Protein structure modeling indicates hexahistidine‐tag interference with enzyme activity

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Publisher
Wiley
Copyright
Copyright © 2008 Wiley Subscription Services
ISSN
0887-3585
eISSN
1097-0134
D.O.I.
10.1002/prot.21905
Publisher site
See Article on Publisher Site

Abstract

Unusual kinetic characteristics of tropinone reductase, an enzyme in the family of short chain dehydrogenases, prompted to investigate a possible impact of the hexahistidine affinity tag on catalytic properties. Comparison of enzymes from Solanum dulcamara, Solanaceae, tagged at the N‐terminus or at the C‐terminus revealed that the C‐terminally tagged form was functionally impaired. Protein modeling indicated that the hexahistidine tag attached at the C‐terminus but not at the N‐terminus of the polypeptide can interfere with the active site by steric or electrostatic interactions. In consequence, protein modeling is suggested before enzyme expression with affinity tags to estimate possible interactions of affinity tags with the active center. Proteins 2008. © 2008 Wiley‐Liss, Inc.

Journal

Proteins: Structure Function and BioinformaticsWiley

Published: Jan 1, 2008

Keywords: ; ; ; ; ; ; ; ;

References

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