PMID: 16636455Apr 26, 2006Paper

Mutational study on alphaGln90 of Fe-type nitrile hydratase from Rhodococcus sp. N771

Bioscience, Biotechnology, and Biochemistry
Hiromi TakaradaMasafumi Odaka

Abstract

Nitrile hydratase (NHase) from Rhodococcus sp. N771 is a non-heme iron enzyme having post-translationally modified cysteine ligands, alphaCys112-SO2H and alphaCys114-SOH. We replaced alphaGln90, which is conserved in all known NHases and involved in the hydrogen-bond network around the catalytic center, with glutamic acid or asparagine. The kcat of alphaQ90E and alphaQ90N mutants decreased to 24% and 5% that of wild type respectively, but the effect of mutations on Km was not very significant. In both mutants, the alphaCys114-SOH modification appeared to be responsible for the catalysis as in native NHase. We crystallized the nitrosylated alphaQ90N mutant and determined its structure at a resolution of 1.43 A. The structure was basically identical to that of native nitrosylated NHase except for the mutated site and its vicinity. The structural difference between native and alphaQ90N mutant NHases suggested the importance of the hydrogen bond networks between alphaGln90 and the iron center for the catalytic activity.

References

Nov 1, 1992·Trends in Biotechnology·M KobayashiH Yamada
Apr 30, 1990·Biochemical and Biophysical Research Communications·T NagamuneI Endo
May 20, 1998·Nature Structural Biology·S NagashimaI Endo
Oct 3, 1998·Acta Crystallographica. Section D, Biological Crystallography·A T BrüngerG L Warren
Apr 11, 2001·Journal of Inorganic Biochemistry·I EndoM Odaka
Jul 4, 2001·Biochemistry·V C PopescuM J Nelson
Nov 9, 2001·Biochemical and Biophysical Research Communications·A MiyanagaT Wakagi
Sep 19, 2002·Journal of the American Chemical Society·Jason ShearerJulie A Kovacs
Sep 12, 2003·Protein Engineering·Dolores Gonzalez-PacanowskaDaniel V Santi
Sep 18, 2003·Journal of the American Chemical Society·Masanari TsujimuraIsao Endo
Sep 25, 2003·Journal of Mass Spectrometry : JMS·Julie M StevensIsabelle Artaud
Nov 26, 2003·Biochemical and Biophysical Research Communications·Shinji HouraiKazunori Yanagi
Nov 26, 2003·The Journal of Biological Chemistry·Akira HirataBunzo Mikami
Jan 14, 2004·European Journal of Biochemistry·Akimasa MiyanagaTakayoshi Wakagi
Jan 28, 2004·Annual Review of Pharmacology and Toxicology·Leslie B PooleAl Claiborne
Jan 1, 1997·Methods in Enzymology·Zbyszek Otwinowski, Wladek Minor

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Citations

Mar 12, 2010·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Yasuaki YamanakaMasafumi Odaka
Aug 6, 2010·Biotechnology Advances·Shreenath Prasad, Tek Chand Bhalla
Sep 30, 2009·Journal of the American Chemical Society·Takatoshi ArakawaMasafumi Odaka
Jan 30, 2015·Protein Science : a Publication of the Protein Society·Heather R BrodkinMary Jo Ondrechen
Feb 13, 2016·The Journal of Biological Chemistry·Micah T NelpVahe Bandarian
Oct 23, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Zhongyi ChengLukasz Peplowski
Mar 24, 2021·International Journal of Biological Macromolecules·Junling GuoZhemin Zhou
Feb 1, 2008·Biophysical Journal·Karina Kubiak, Wieslaw Nowak
May 15, 2007·Inorganic Chemistry·Kathrin H HopmannFahmi Himo

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