Bookedited by Karen N. Allen.
Summary: High-energy phosphate ester bonds are often thought of as the energetic currency of the cell. Phosphoryl transfer is involved in processes ranging from the biosynthesis and catabolism of carbohydrates and polysaccharides, nucleic acids, and cofactors, to the regulation if cellular pathways. In addition, Phosphoryl groups on small-molecule metabolites confer important properties - providing excellent "handles" for enzyme bonding, trapping metabolic intermediates within the cell, and controlling the reversibility of metabolic pathways. The phosphorylation states of proteins also afford key control points for signal transduction and the regulation of myriad critical physiological processes. For this reason, kinases are extremely important therapeutic targets and there is significant interest in developing diverse approaches to regulate their function through small-molecule inhibitors. We present here the less studied but equally important complement of phosphate ester chemistry, namely the coordinated action of phosphatases. Phosphatases represent control points in pathways and new opportunities for use in chemoenzymatic synthesis. In many cases, such critical players may be functionally unannotated. However, there are many challenges in their study - spurring on the efforts described in this volume ... This volume of Methods in Enzymology on phosphatases which draws upon approaches from the varied disciplines of computational and organic chemistry, biophysics, biochemistry, bioinformatics, and cell biology to address the structure, mechanism, and regulation of phosphatases."--Preface.
Contents:
Empirical Valence Bond Simulations of Organophosphate Hydrolysis: Theory and Practice / Miha Purg and Shina Caroline Lynn Kamerlin
Analysis of Phosphoryl-Transfer Enzymes With QM/MM Free Energy Simulations / Daniel Roston, Xiya Lu, Dong Fang, Darren Demapan and Qiang Cui
Defining Dynamics of Membrane-Bound Pyrophosphatases by Experimental and Computational Single-Molecule FRET / Steven P.D. Harborne, Jannik Strauss, Ainoleena Turku, Matthew A. Watson, Roman Tuma, Sarah A. Harris and Adrian Goldman
A Simple Strategy to Determine the Dependence of Membrane-Bound Pyrophosphatases on K+ as a Cofactor / Jannik Strauss, Craig Wilkinson, Keni Vidilaseris, Steven P.D. Harborne and Adrian Goldman
Crystallization of Liganded Phosphatases in the HAD Superfamily / Christine Harvey, Katherine H. O'Toole and Karen N. Allen
Structure, Mechanism, and Substrate Profiles of the Trinuclear Metallophosphatases From the Amidohydrolase Superfamily / Swapnil V. Ghodge and Frank M. Raushel
Affinities and Comparisons of Enzyme States by Principal Component Analysis of NMR Spectra, Automated Using TREND Software / Jia Xu and Steven R. Van Doren
Assessment and Impacts of Phosphorylation on Protein Flexibility of the [alpha]-D-Phosphohexomutases / Kyle M. Stiers and Lesa J. Beamer
Chemical Tools for Studying the Impact of cis/trans Prolyl Isomerization on Signaling: A Case Study on RNA Polymerase II Phosphatase Activity and Specificity / Nathaniel Tate Burkholder, Brenda Medellin, Seema Irani, Wendy Matthews, Scott A. Showalter and Yan Jessie Zhang
Quantitative Analysis of Intracellular Response Regulator Phosphatase Activity of Histidine Kinases / Rong Gao and Ann M. Stock
Measuring the Activities of Two-Component Regulatory System Phosphatases / Robert B. Bourret and Ruth E. Silversmith
Assembly of PGAM5 Into Multimeric Complexes Provides a Mechanism for Allosteric Regulation of Phosphatase Activity / Peter Tipton, Tong Su and Mark Hannink
Purification of Lipin and Measurement of Phosphatidic Acid Phosphatase Activity From Liposomes / Mitchell E. Granade and Thurl E. Harris
Combining SAMDI Mass Spectrometry and Peptide Arrays to Profile Phosphatase Activities / Lindsey C. Szymczak, Che-Fan Huang, Eric J. Berns and Milan Mrksich
Protein Chemical Approaches to Understanding PTEN Lipid Phosphatase Regulation / Daniel R. Dempsey and Philip A. Cole.