Determination of molecular weight of the protein moiety in protein-detergent complexes without direct knowledge of detergent binding

Abstract

Sedimentation equilibrium measurements can be used to determine the molecular weight of the protein moiety of a protein-detergent complex without prior knowledge of detergent binding. The procedure is to adjust the solvent density by addition of D2O so as to blank out the contribution of bound detergent to the sedimentation potential. An approximate measure of detergent binding can be obtained from the effect of solvent density on the sedimentation result. The procedure is also applicable to protein-lipid complexes. It can be used for complexes containing both lipid and detergent if the lipid content is known. The use of the method is demonstrated by experimental data for the AI polypeptide of serum high density lipoprotein, in separate complexes with nonionic detergents and with a phospholipid.