Author : Venkatramanan Krishnamani
Publisher :
ISBN 13 : 9781124208350
Total Pages : 120 pages
Book Rating : 4.2/5 (83 download)
Book Synopsis Structure of Partially Denatured Bacteriorhodopsin in Detergent Micelles and Structural Changes During Its Refolding by : Venkatramanan Krishnamani
Download or read book Structure of Partially Denatured Bacteriorhodopsin in Detergent Micelles and Structural Changes During Its Refolding written by Venkatramanan Krishnamani and published by . This book was released on 2010 with total page 120 pages. Available in PDF, EPUB and Kindle. Book excerpt: The partially denatured state of bacteriorhodopsin (bR), a model hepta-helical integral membrane protein that acts as a proton pump in the membranes of Halobacterium salinarum, in SDS micelles was studied to understand the sequence dependent structural perturbations. Two-frequency pulsed electron paramagnetic resonance (EPR) (double electron-electron spin resonance, DEER) investigation combined with molecular dynamics (MD) of denaturation of individual helices of bR was used in an attempt to unravel the structural details of bR in sodium dodecyl sulfate (SDS) micelles. The results from the DEER experiments reveal a discrete heterogeneous ensemble of structures of bR in the detergent. The MD simulations show the role of charged residues in unfolding and the partitioning preference of the helices in SDS micelles. The refolding kinetics from this partially denatured starting state to a functional regenerated state using stopped flow continuous wave EPR measurements revealed that the secondary structure formation is faster than 140 msec and further helix association events followed a two step process. Majority of the structural changes happen before the functional recovery of the protein in the first step with a time constant of 3-5 sec followed by minor structural changes occurring in concert with the retinal chromophore recovery with varying association time constants for each helix pair.