Observation of persistent α‐helical content and discrete types of backbone disorder during a molten globule to ordered peptide transition via deep‐UV resonance Raman spectroscopy. (1st June 2013)

• Record Type: Journal Article
• Authors: Brown, Mia C.; Mutter, Andrew C.; Koder, Ronald L.; JiJi, Renee D.; Cooley, Jason W.
• Is Part Of: Journal of Raman spectroscopy. Volume 44:Number 7(2013:Jul.)
• Page Start: 957
• Page End: 962
• Publication Date: 2013-06-01
• Subjects: Raman spectroscopy -- Periodicals; 535.846
• DOI: 10.1002/jrs.4316
• Languages: English
• ISSNs: 0377-0486

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p>The molten globule (MG) state can aid in the folding of a protein to a functional structure and is loosely defined as an increase in structural disorder with conservation of the ensemble secondary structure content. Simultaneous observation of persistent secondary structure content with increased disorder has remained experimentally problematic. As a consequence, modeling how the MG state remains stable and how it facilitates proper folding remains difficult due to a lack of amenable spectroscopic techniques to characterize this class of partially unfolded proteins. Previously, deep‐UV resonance Raman (dUVRR) spectroscopy has proven useful in the resolution of global and local structural fluctuations in the secondary structure of proteins. In this work, dUVRR was employed to study the MG to ordered transition of a model four‐helix bundle protein, HP7. Both the average ensemble secondary structure and types of local disorder were monitored, without perturbation of the solvent, pH, or temperature. The MG to ordered transition is induced by stepwise coordination of two heme molecules. Persistent dUVRR spectral features in the amide III region at 1295–1301 and 1335–1338 cm<sup>−1</sup> confirm previous observations that HP7 remains predominantly helical in the MG <italic>versus</italic> the fully ordered state. Additionally, these spectra represent the first demonstration of conserved helical content in a MG protein. With successive heme binding, significant losses are observed in the spectral intensity of the amide III<sub>3</sub> and S regions (1230–1260 and 1390 cm<sup>−1</sup>, respectively), which are known to be sensitive to local disorder. These observations indicate that there is a decrease in the structural populations able to explore various extended conformations with successive heme binding events. DUVRR spectra indicate that the first heme coordination between two helical segments diminishes exploration of more elongated backbone structural conformations in the inter‐helical regions. A second heme coordination by the remaining two helices further restricts protein motion. Copyright © 2013 John Wiley & Sons, Ltd.</p> </abstract>