Determination of the folding of proteins as a function of denaturants, osmolytes or ligands using circular dichroism.
Circular dichroism (CD) is an excellent tool for examining the interactions and stability of proteins. This protocol covers methods to obtain and analyze circular dichroism spectra to measure changes in the folding of proteins as a function of denaturants, osmolytes or ligands. Applications include determination of the free energy of folding of a protein, the effects of mutations on protein stability and the estimation of binding constants for the interactions of proteins with other proteins, DNA or ligands, such as substrates or inhibitors. The experiments require 2-5 h.
Greenfield
Norma J
NJ
Department of Neuroscience and Cell Biology, Robert Wood Johnson Medical School, 675 Hoes Lane West, Piscataway, New Jersey 08854-8021, USA.
D002942:Circular Dichroism / Q000379:methods*; D008024:Ligands; D009994:Osmolar Concentration; D011489:Protein Denaturation; D017510:Protein Folding
D008024:Ligands