PMID: 11902881Mar 21, 2002Paper

Measurement of electrostatic interactions in protein folding with the use of protein charge ladders

Journal of the American Chemical Society
R S Negin, Jeffrey D Carbeck

Abstract

This paper describes a new method for the measurement of the role of interactions between charged groups on the energetics of protein folding. This method uses capillary electrophoresis (CE) and protein charge ladders (mixtures of protein derivatives that differ incrementally in number of charged groups) to measure, in a single set of electrophoresis experiments, the free energy of unfolding (DeltaG(D-N)) of alpha-lactalbumin (alpha-LA) as a function of net charge. These same data also yield the hydrodynamic radius, R(H), and net charge measured by CE, Z(CE), of the folded and denatured proteins. Alpha-LA unfolds to a compact denatured state under mildly alkaline conditions; a small increase in R(H) (11%, 2 A) coincides with a large increase in Z(CE) (71%, -4 charge units), relative to the folded state. The increase in Z(CE), in turn, predicts a large pH dependence of free energy of unfolding (-22 kJ/mol per unit increase in pH), due to differences in proton binding in the folded and denatured states. The free energy of unfolding correlates with the square of net charge of the members of the charge ladder. The differential dependence of DeltaG(D-N) on net charge for holo-alpha-LA, (partial differential) DeltaG(D-N)/(partial dif...Continue Reading

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Citations

Oct 2, 2007·Protein Science : a Publication of the Protein Society·Ishara A MillsJonathan A King
Sep 6, 2012·Biotechnology and Bioengineering·Guillaume RousselCatherine Michaux
May 21, 2016·Journal of Peptide Science : an Official Publication of the European Peptide Society·C MichauxE A Perpète
Jun 1, 2007·Electrophoresis·Maria V PiaggioJulio A Deiber
Apr 30, 2005·Electrophoresis·Upma Sharma, Jeffrey D Carbeck
Jan 23, 2003·Protein Science : a Publication of the Protein Society·Tamas BlandlNicholas J Skelton
Aug 15, 2009·Langmuir : the ACS Journal of Surfaces and Colloids·Wai Keen ChungSteven M Cramer
Sep 24, 2004·Journal of the American Chemical Society·Thomas F O'ConnorJeffrey D Carbeck

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