Protein structure modeling indicates hexahistidine-tag interference with enzyme activity

Proteins
Anna-Carolin FreydankBirgit Dräger

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.

References

Nov 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·S Henikoff, J G Henikoff
Apr 1, 1992·Anales de medicina interna : organo oficial de la Sociedad Española de Medicina Interna·B Pinilla LlorenteJ García Castaño
Apr 1, 1968·Experimental Cell Research·O L GamborgK Ojima
Sep 1, 1993·Proteins·S Henikoff, J G Henikoff
Oct 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·K NakajimaY Yamada
Jun 6, 1998·Proceedings of the National Academy of Sciences of the United States of America·K NakajimaY Yamada
Jun 2, 2000·Biochemical and Biophysical Research Communications·A P AraújoL M Beltramini
Feb 27, 2003·Chemico-biological Interactions·Udo OppermannHans Jörnvall
May 7, 2003·Current Opinion in Biotechnology·Takashi Hashimoto, Yasuyuki Yamada
Sep 10, 2003·Protein Expression and Purification·Richard J JennyRoger L Lundblad
Mar 26, 2004·Natural Product Reports·Birgit Dräger
Apr 28, 2004·Bioorganic & Medicinal Chemistry·Anja BayerJoachim Stöckigt
Sep 1, 2004·Trends in Plant Science·Kirsi-Marja Oksman-Caldentey, Dirk Inzé
Sep 24, 2004·Nature·Anthony G MillgatePhilip J Larkin
Jan 12, 2005·Journal of Experimental Botany·Ute RichterBirgit Dräger
May 24, 2005·Phytochemistry·Andrea BrockBirgit Dräger
Jan 24, 2006·Protein Expression and Purification·José ArnauJohn Pedersen
Jun 20, 2006·Proceedings of the National Academy of Sciences of the United States of America·Takao KoedukaEran Pichersky
Aug 15, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Mouna Amor-MahjoubMoncef Ladjimi
Oct 20, 2006·Proceedings of the National Academy of Sciences of the United States of America·Ling QinShelagh Ferguson-Miller
Jan 6, 2007·The Journal of Biological Chemistry·Hiromichi MinamiFumihiko Sato
Feb 16, 2007·Journal of Molecular Biology·Yoshiaki YasutakeTomohiro Tamura
Mar 1, 2007·Acta Crystallographica. Section D, Biological Crystallography·Mike CarsonLawrence J Delucas
Dec 19, 2007·The Alkaloids. Chemistry and Biology·Stefan Biastoff, Birgit Dräger
Apr 30, 1998·The Journal of Physical Chemistry. B·A D MacKerellM Karplus

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Citations

Jul 14, 2010·Journal of Molecular Modeling·Ying-Wu LinLi-Fu Liao
Jan 1, 1989·Archives of Gerontology and Geriatrics·S DamjanovichB Somogyi
Sep 1, 1991·Archives of Gerontology and Geriatrics·I Zs -Nagy, R A Floyd
May 31, 2011·Biochemistry·Megan C ThielgesMichael D Fayer
Jun 8, 2011·Antimicrobial Agents and Chemotherapy·Hyun KimYasuhiko Suzuki
Feb 10, 2009·Applied and Environmental Microbiology·Hanna KontkanenJohanna Buchert
Nov 22, 2011·Biochemistry. Biokhimii︠a︡·Yu A ModestovaN N Ugarova
Aug 22, 2013·Plant Molecular Biology·Thomas De MeyerAnn Depicker
Nov 15, 2011·Biotechnology Journal·Farhat KhanEvelina Angov
Nov 3, 2010·Mass Spectrometry Reviews·Ganesh Kumar AgrawalRandeep Rakwal
Mar 16, 2011·Protein Science : a Publication of the Protein Society·Marcia R MaukA Grant Mauk
May 23, 2015·The Journal of Physical Chemistry. B·Paolo A CalligariGerald R Kneller
Dec 14, 2012·PloS One·David P SargeantMartin R Schiller
Nov 29, 2017·Organic & Biomolecular Chemistry·Julian HaasSteffen Lüdeke
Nov 16, 2018·Journal of Medical Virology·Muhammad Usman MirzaMuhammad Idrees
Aug 23, 2020·Biomolecules·Emily I SparkesLu Shin Wong
Feb 3, 2021·Journal of Reproductive Immunology·Jacqueline GuettlerMartin Gauster
Aug 17, 2017·Biochemistry·Avinash Kumar ThakurLiviu Movileanu
Aug 22, 2021·Biotechnology Advances·Hèctor López-LagunaUgutz Unzueta

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