Diamond Publications - Search Results

Search Again...

These results include publications that have not yet been reviewed and processed by Diamond Light Source. To view only processed publications click here.

You can use the folder icon under Actions to collate papers as required. You can then click on View Folder in the left-hand navigation to review and/or download your folder.

One item found (0 items not approved).

Instruments / Technical Area	Publication Details	Published	Actions
B23-Circular Dichroism	Purification of bacterial membrane sensor kinases and biophysical methods for determination of their ligand and inhibitor interactions R. Hussain , S. E. Harding , C. S. Hughes , P. Ma , S. G. Patching , S. Edara , G. Siligardi , P. J. F. Henderson , M. K. Phillips-jones Biochemical Society Transactions 44, 810 - 823 DOI: 10.1042/BST20160023 Diamond Proposal Number(s): [12003, 12406, 12310] Open Access Show Abstract ▼ Abstract: This article reviews current methods for the reliable heterologous overexpression in Escherichia coli and purification of milligram quantities of bacterial membrane sensor kinase (MSK) proteins belonging to the two-component signal transduction family of integral membrane proteins. Many of these methods were developed at Leeds alongside Professor Steve Baldwin to whom this review is dedicated. It also reviews two biophysical methods that we have adapted successfully for studies of purified MSKs and other membrane proteins–synchrotron radiation circular dichroism (SRCD) spectroscopy and analytical ultracentrifugation (AUC), both of which are non-immobilization and matrix-free methods that require no labelling strategies. Other techniques such as isothermal titration calorimetry (ITC) also share these features but generally require high concentrations of material. In common with many other biophysical techniques, both of these biophysical methods provide information regarding membrane protein conformation, oligomerization state and ligand binding, but they possess the additional advantage of providing direct assessments of whether ligand binding interactions are accompanied by conformational changes. Therefore, both methods provide a powerful means by which to identify and characterize inhibitor binding and any associated protein conformational changes, thereby contributing valuable information for future drug intervention strategies directed towards bacterial MSKs.	Jun 2016

Instruments / Technical Area

Publication Details

Published

Actions

B23-Circular Dichroism

Purification of bacterial membrane sensor kinases and biophysical methods for determination of their ligand and inhibitor interactions

R. Hussain , S. E. Harding , C. S. Hughes , P. Ma , S. G. Patching , S. Edara , G. Siligardi , P. J. F. Henderson , M. K. Phillips-jones

Biochemical Society Transactions 44, 810 - 823

DOI: 10.1042/BST20160023

Diamond Proposal Number(s): [12003, 12406, 12310]