Coordination sphere of the third metal site is essential to the activity and metal selectivity of alkaline phosphatases.

Protein Science : a Publication of the Protein Society
Dimitris KoutsioulisPirkko Heikinheimo

Abstract

Alkaline phosphatases (APs) are commercially applied enzymes that catalyze the hydrolysis of phosphate monoesters by a reaction involving three active site metal ions. We have previously identified H135 as the key residue for controlling activity of the psychrophilic TAB5 AP (TAP). In this article, we describe three X-ray crystallographic structures on TAP variants H135E and H135D in complex with a variety of metal ions. The structural analysis is supported by thermodynamic and kinetic data. The AP catalysis essentially requires octahedral coordination in the M3 site, but stability is adjusted with the conformational freedom of the metal ion. Comparison with the mesophilic Escherichia coli, AP shows differences in the charge transfer network in providing the chemically optimal metal combination for catalysis. Our results provide explanation why the TAB5 and E. coli APs respond in an opposite way to mutagenesis in their active sites. They provide a lesson on chemical fine tuning and the importance of the second coordination sphere in defining metal specificity in enzymes. Understanding the framework of AP catalysis is essential in the efforts to design even more powerful tools for modern biotechnology.

References

Mar 20, 1991·Journal of Molecular Biology·E E Kim, H W Wyckoff
Dec 15, 1996·Structure·P HeikinheimoA Goldman
Feb 15, 2000·European Journal of Biochemistry·M RinaV Bouriotis
Jun 30, 2000·Journal of Molecular Biology·B StecE R Kantrowitz
Mar 15, 2001·Proceedings of the National Academy of Sciences of the United States of America·P HeikinheimoA Goldman
Oct 9, 2001·European Journal of Biochemistry·I TsigosV Bouriotis
Feb 6, 2002·Chembiochem : a European Journal of Chemical Biology·B H MullerJ C Boulain
Mar 23, 2002·Protein Science : a Publication of the Protein Society·Cheryl L WojciechowskiEvan R Kantrowitz
Jun 27, 2002·Journal of Molecular Biology·Maaike de BackerEdward Hough
Oct 24, 2002·Acta Crystallographica. Section D, Biological Crystallography·Paul D AdamsThomas C Terwilliger
Oct 26, 2002·The Journal of Biological Chemistry·Cheryl L Wojciechowski, Evan R Kantrowitz
Jan 4, 2003·The Journal of Biological Chemistry·Salvino D'AmicoGeorges Feller
Sep 1, 1994·Acta Crystallographica. Section D, Biological Crystallography·UNKNOWN Collaborative Computational Project, Number 4
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
Apr 5, 2005·Acta Crystallographica. Section D, Biological Crystallography·Airlie J McCoyRandy J Read
Jun 11, 2005·Journal of Molecular Biology·Paola LlinasMarie Hélène Le Du
Mar 23, 2006·Acta Crystallographica. Section D, Biological Crystallography·Jay Painter, Ethan A Merritt
Jan 3, 2007·Journal of Molecular Biology·Ellen WangPirkko Heikinheimo
Jan 31, 2007·Biochemistry·Esko OksanenPirkko Heikinheimo
Mar 8, 2008·Acta Crystallographica. Section D, Biological Crystallography·Ivan DokmanićSanja Tomić
Apr 16, 2008·Protein Engineering, Design & Selection : PEDS·Dimitris KoutsioulisVassilis Bouriotis
May 9, 2008·Journal of Molecular Biology·Yu-Ming Lee, Carmay Lim
Nov 4, 2008·Biochimica Et Biophysica Acta·Ronny HellandBjarni Asgeirsson

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Citations

Mar 7, 2014·Acta Crystallographica. Section D, Biological Crystallography·Shigeki AraiRyota Kuroki
Feb 7, 2014·Journal of the American Chemical Society·Edina RostaGerhard Hummer
Jun 21, 2015·Journal of the American Chemical Society·Alexandre BarrozoShina C L Kamerlin
Nov 13, 2010·Metallomics : Integrated Biometal Science·Cynthia M Dupureur

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