Hierarchy in guanidine unfolding of DLC8 dimer: regulatory functional implications

Krishna Mohan, P. M. ; Joshi, Mamata V. ; Hosur, Ramakrishna V. (2009) Hierarchy in guanidine unfolding of DLC8 dimer: regulatory functional implications Biochimie, 91 (3). pp. 401-407. ISSN 0300-9084

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Official URL: http://linkinghub.elsevier.com/retrieve/pii/S03009...

Related URL: http://dx.doi.org/10.1016/j.biochi.2008.10.013


Folding-unfolding caused by environmental changes play crucial regulatory roles in protein functions. To gain an insight into these for DLC8, a cargo adaptor in dynein motor complex, we investigated here the unfolding of homodimeric DLC8 by GdnHCl, a standard unfolding agent. Fluorescence spectroscopy revealed a three-state unfolding transition with midpoints at 1.5 and 4.0 M GdnHCl. The HSQC spectrum at 1.5 M GdnHCl displayed peaks belonging to a folded monomer. NMR chemical shift perturbations, line broadening effects and 15N relaxation measurements at low GdnHCl concentrations identified a hierarchy in the unfolding process, with the dimer interface - the cargo binding site - being the most susceptible followed by the helices in the interior. Similar observations were made earlier for small pH perturbations and thus the early unfolding events appear to be intrinsic to the protein. These, by virtue of their location, influence target binding efficacies and thus have important regulatory implications.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:Dynein Light Chain Protein; Nuclear Magnetic Resonance; Circular Dichroism; Fluorescence Spectroscopy; Protein Folding; Monomeric Intermediate
ID Code:16605
Deposited On:15 Nov 2010 13:35
Last Modified:03 Jun 2011 08:28

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