Histidine Phosphorylation, 1st ed. 2020 Methods and Protocols Methods in Molecular Biology Series, Vol. 2077

Thisvolumedetails 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. 

1. Protein Dynamics in Phosphoryl-transfer Signaling Mediated by Two-component Systems

Felipe Trajtenbergand Alejandro Buschiazzo

2. A High-throughput Strategy for Recombinant Protein Expression and Solubility Screening in Escherichia coli – A Case of Sensor Histidine Kinase

Agnieszka Szmitkowska, Blanka Pekárová, and Jan Hejátko

3. SDS-PAGE and Dot Blot Autoradiography: Tools for Quantifying Histidine Kinase Autophosphorylation

Jonathan T. Fischer, Ilana Heckler, and Elizabeth M. Boon

4. A Quantitative Method for the Measurement of Protein Histidine Phosphorylation

Paul V. Attwood

5. Analysis of 1- and 3-phosphohistidine (pHis) Protein Modification using Model Enzymes Expressed in Bacteria

Alice K. M. Clubbs Coldron, Dominic P. Byrne, and Patrick A. Eyers

6. Determination of Phosphohistidine Stoichiometry in Histidine Kinases by Intact Mass Spectrometry

Lauren J. Tomlinson, Alice K. M. Clubbs Coldron, Patrick A. Eyers, and Claire E. Eyers

8. In Vitro Assays for Measuring Protein Histidine Phosphatase Activity

Brandon S. McCullough and Amy M. Barrios

9. Structural and Functional Characterization of Autophosphorylation in Bacterial Histidine Kinases

Laura Miguel-Romero, Cristina Mideros-Mora, Alberto Marina, and Patricia Casino

10. Manipulation of Bacterial Signalling using Engineered Histidine Kinases

Kimberly A. Kowallis, Samuel W. Duvall, Wei Zhao, and W. Seth Childers

11. Development of a Light-Dependent Protein Histidine Kinase

Aleksandra Bury and Klaas J. Hellingwerf

12. Empirical Evidence of Cellular Histidine Phosphorylation by Immunoblotting using pHis mAbs

Rajasree Kalagiri, Kevin Adam, and Tony Hunter

13. Immunohistochemistry (IHC):  Chromogenic Detection of 3-Phosphohistidine Proteins in Formaldehyde-Fixed, Frozen Mouse Liver Tissue Sections

14. Subcellular Localisation of Histidine Phosphorylated Proteins through Indirect Immunofluorescence

Kevin Adam and Tony Hunter

15. High throughput Characterisation of Histidine Phosphorylation sites using UPAX and Tandem Mass Spectrometry

Gemma Hardman and Claire E. Eyers

16. Proteome Bioinformatics Methods for Studying Histidine Phosphorylation

Andrew R Jones and Oscar Martin Camacho

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts