A potential role for the analytical ultracentrifuge in the experimental measurement of protein valence.
A method is described whereby sedimentation velocity is combined with equilibrium dialysis to determine the net charge (valence) of a protein by using chromate as an indicator ion for assessing the extent of the Donnan redistribution of small ions. The procedure has been used in experiments on bovine serum albumin under slightly alkaline conditions (pH 8.0, I 0.05) to illustrate its application to a system in which the indicator ion and protein both bear net negative charge and on lysozyme under slightly acidic conditions (pH 5.0, I 0.10) to illustrate the situation where chromate is a counterion.
Winzor
D J
DJ
Department of Biochemistry, School of Molecular and Microbial Sciences, University of Queensland, Brisbane, Queensland 4072, Australia. winzor@biosci.uq.edu.au
Carrington
L E
LE
Harding
S E
SE
D000818:Animals; D002417:Cattle; D002840:Chromates / Q000737:chemistry; D003956:Dialysis; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000737:chemistry; D009113:Muramidase / Q000032:analysis*; D012710:Serum Albumin, Bovine / Q000032:analysis*; D055672:Static Electricity; D014461:Ultracentrifugation / Q000379:methods*
D002840:Chromates; D007202:Indicators and Reagents; D012710:Serum Albumin, Bovine; D009113:Muramidase