Weak and Transient Protein Interactions Determined by Solid‐State NMR. Issue 23 (21st April 2016)

• Record Type: Journal Article
• Authors: Dannatt, Hugh R. W.; Felletti, Michele; Jehle, Stefan; Wang, Yao; Emsley, Lyndon; Dixon, Nicholas E.; Lesage, Anne; Pintacuda, Guido
• Is Part Of: Angewandte Chemie international edition. Volume 55:Issue 23(2016)
• Page Start: 6638
• Page End: 6641
• Publication Date: 2016-04-21
• Subjects: DNA replication -- magic angle spinning -- solid-state NMR -- protein structure -- protein–protein interactions; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201511609
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Despite their roles in controlling many cellular processes, weak and transient interactions between large structured macromolecules and disordered protein segments cannot currently be characterized at atomic resolution by X‐ray crystallography or solution NMR. Solid‐state NMR does not suffer from the molecular size limitations affecting solution NMR, and it can be applied to molecules in different aggregation states, including non‐crystalline precipitates and sediments. A solid‐state NMR approach based on high magnetic fields, fast magic‐angle sample spinning, and deuteration provides chemical‐shift and relaxation mapping that enabled the characterization of the structure and dynamics of the transient association between two regions in an 80 kDa protein assembly. This led to direct verification of a mechanism of regulation of E. coli DNA metabolism. Abstract : In a spin : A solid‐state NMR approach based on high magnetic fields, fast magic‐angle spinning, and deuteration was used to provide chemical‐shift and relaxation mapping for characterizing the transient association between two regions in a 80 kDa protein assembly, the homotetrameric ssDNA‐binding protein (SSB). Comparison of the wildtype (wt) and the truncated mutant SSBΔCt led to direct verification of a mechanism of regulation of E. coli DNA metabolism.