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Protein structure modeling indicates hexahistidine‐tag interference with enzyme activity
Author(s) -
Freydank AnnaCarolin,
Brandt Wolfgang,
Dräger Birgit
Publication year - 2008
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.21905
Subject(s) - enzyme , biochemistry , steric effects , chemistry , active site , stereochemistry , affinity chromatography , n terminus , peptide sequence , gene
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.