Dynamics of ribonuclease A and ribonuclease S: Computational and experimental studies

Nadig, Gautham and Ratnaparkhi, Girish S and Varadarajan, Raghavan and Vishveshwara, Saraswathi (1996) Dynamics of ribonuclease A and ribonuclease S: Computational and experimental studies. In: Protein Science, 05 (10). pp. 2104-2114.

PDF Dynamics_of_ribonuclease.pdf - Published Version Restricted to Registered users only Download (3MB) | Request a copy

Abstract

RNase S is a complex consisting of two proteolytic fragments of RNase A: the S peptide (residues 1-20) and S protein (residues 21-124). RNase S and RNase A have very similar X-ray structures and enzymatic activities. previous experiments have shown increased rates of hydrogen exchange and greater sensitivity to tryptic cleavage for RNase S relative to RNase A. It has therefore been asserted that the RNase S complex is considerably more dynamically flexible than RNase A. In the present study we examine the differences in the dynamics of RNaseS and RNase A computationally, by MD simulations, and experimentally, using trypsin cleavage as a probe of dynamics. The fluctuations around the average solution structure during the simulation were analyzed by measuring the RMS deviation in coordinates. No significant differences between RNase S and RNase A dynamics were observed in the simulations. We were able to account for the apparent discrepancy between simulation and experiment by a simple model, According to this model, the experimentally observed differences in dynamics can be quantitatively explained by the small amounts of free S peptide and S protein that are present in equilibrium with the RNase S complex. Thus, folded RNase A and the RNase S complex have identical dynamic behavior, despite the presence of a break in polypeptide chain between residues 20 and 21 in the latter molecule. This is in contrast to what has been widely believed for over 30 years about this important fragment complementation system.

Item Type:	Journal Article
Publication:	Protein Science
Publisher:	Cold Spring Harbor Laboratory Press
Additional Information:	Copyright of this article belongs to Cold Spring Harbor Laboratory Press.
Department/Centre:	Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited:	09 Jul 2009 10:28
Last Modified:	19 Sep 2010 05:28
URI:	http://eprints.iisc.ac.in/id/eprint/19401

Actions (login required)

View Item