The energetics of barstar denaturation have been studied by CD and scanning microcalorimetry in an extended range of pH and salt concentration. It was shown that, upon increasing temperature, barstar undergoes a transition to the denatured state that is well approximated by a two-state transition in solutions of high ionic strength. This transition is accompanied by significant heat absorption and an increase in heat capacity. The denaturational heat capacity increment at approximately 75 degrees C was found to be 5.6 +/- 0.3 kJ K-1 mol-1. In all cases, the value of the measured enthalpy of denaturation was notably lower than those observed for other small globular proteins. In order to explain this observation, the relative contributions of hydration and the disruption of internal interactions to the total enthalpy and entropy of unfolding were calculated. The enthalpy and entropy of hydration were found to be in good agreement with those calculated for other proteins, but the enthalpy and entropy of breaking internal interactions were found to be among the lowest for all globular proteins that have been studied. Additionally, the partial specific heat capacity of barstar in the native state was found to be 0.37 +/- 0.03 cal K-1 g-1, which is higher than what is observed for most globular proteins and suggests significant flexibility in the native state. It is known from structural data that barstar undergoes a conformational change upon binding to its natural substrate barnase.(ABSTRACT TRUNCATED AT 250 WORDS) Study holds ProTherm entries: 2577, 2578, 2579 Extra Details: barstar; protein stability; scanning microcalorimetry
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:19 p.m.
|Number of data points||11|
|Proteins||Barstar ; Barstar|
|Assays/Quantities/Protocols||Experimental Assay: dCp pH:9.5 ; Experimental Assay: dHcal pH:9.5 ; Experimental Assay: Tm pH:9.5 ; Experimental Assay: dHvH pH:9.5 ; Experimental Assay: dCp pH:8.0 ; Experimental Assay: dHcal pH:8.0 ; Experimental Assay: Tm pH:8.0 ; Experimental Assay: dHvH pH:8.0 ; Experimental Assay: dHcal pH:6.3 ; Experimental Assay: Tm pH:6.3 ; Experimental Assay: dHvH pH:6.3|
|Libraries||Mutations for sequence KKAVINGEQIRSISDLHQTLKKELALPEYYGENLDALWDCLTGWVEYPLVLEWRQFEQSKQLTENGAESVLQVFREAKAEGCDITIILS|