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This volume details the current understanding of roles and regulation on histidine phosphorylation, describing methods for the characterization of protein phosphorylation on histidine. Chapters guide readers through in vitro systems, cell-based systems, comprehensive background review articles on histidine kinases and phosphatases. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Histidine Phosphorylation: Methods and Protocols aims to ensure successful results in the further study of this rapidly growing field.
List of contents
Protein Dynamics in Phosphoryl-transfer Signaling Mediated by Two-component Systems.- A High-throughput Strategy for Recombinant Protein Expression and Solubility Screening in Escherichia coli - A Case of Sensor Histidine Kinase.- SDS-PAGE and Dot Blot Autoradiography: Tools for Quantifying Histidine Kinase Autophosphorylation.- A Quantitative Method for the Measurement of Protein Histidine Phosphorylation.- Analysis of 1- and 3-phosphohistidine (pHis) Protein Modification using Model Enzymes Expressed in Bacteria.- Determination of Phosphohistidine Stoichiometry in Histidine Kinases by Intact Mass Spectrometry.- Protein Phosphohistidine Phosphatases of the HP Superfamily.- In Vitro Assays for Measuring Protein Histidine Phosphatase Activity.- Structural and Functional Characterization of Autophosphorylation in Bacterial Histidine Kinases.- Manipulation of Bacterial Signalling using Engineered Histidine Kinases.- Development of a Light-Dependent Protein Histidine Kinase.- Empirical Evidence of Cellular Histidine Phosphorylation by Immunoblotting using pHis mAbs.- Immunohistochemistry (IHC): Chromogenic Detection of 3-Phosphohistidine Proteins in Formaldehyde-Fixed, Frozen Mouse Liver Tissue Sections.- Subcellular Localisation of Histidine Phosphorylated Proteins through Indirect Immunofluorescence.- High throughput Characterisation of Histidine Phosphorylation sites using UPAX and Tandem Mass Spectrometry.- Proteome Bioinformatics Methods for Studying Histidine Phosphorylation.
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