1s02
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(New page: 200px<br /><applet load="1s02" size="450" color="white" frame="true" align="right" spinBox="true" caption="1s02, resolution 1.9Å" /> '''EFFECTS OF ENGINEERED...)
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Revision as of 23:52, 20 November 2007
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EFFECTS OF ENGINEERED SALT BRIDGES ON THE STABILITY OF SUBTILISIN BPN'
Overview
Variants designed using PROTEUS have been produced in an attempt to, engineer stabilizing salt bridges into subtilisin BPN'. All the mutants, constructed by site-directed mutagenesis were secreted by Bacillus, subtilis, except L75K. Q19E, expressed as a single variant and also in a, double variant, Q19E/Q271E, appears to form a stabilizing salt bridge, based on X-ray crystal structure determination and differential scanning, calorimeter measurements. Although the double mutant was found to be less, thermodynamically stable than the wild-type, it did exhibit an autolytic, stability about two-fold greater under hydrophobic conditions. Four, variants, A98K, S89E, V26R and L235R, were found to be nearly identical to, wild-type in thermal stability, indicative of stable structures without, evidence of salt bridge formation. Variants Q271E, V51K and T164R led to, structures that resulted in varying degrees of thermodynamic and autolytic, instability. A computer-modeling analysis of the PROTEUS predictions, reveals that the low percentage of salt bridge formation is probably due, to an overly simplistic electrostatic model, which does not account for, the geometry of the pairwise interactions.
About this Structure
1S02 is a Single protein structure of sequence from Bacillus amyloliquefaciens with CA and SO4 as ligands. Active as Subtilisin, with EC number 3.4.21.62 Full crystallographic information is available from OCA.
Reference
Effects of engineered salt bridges on the stability of subtilisin BPN'., Erwin CR, Barnett BL, Oliver JD, Sullivan JF, Protein Eng. 1990 Oct;4(1):87-97. PMID:2127106
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