Sarfo Kwabena J, Winstone Tara L, Papish Andriyka L, Howell Jenika M, Kadir Hakan, Vogel Hans J, Turner Raymond J
Department of Biological Sciences, University of Calgary, 2500 University Dr NW, Calgary, Alta., Canada T2N 1N4.
Biochem Biophys Res Commun. 2004 Mar 5;315(2):397-403. doi: 10.1016/j.bbrc.2004.01.070.
Escherichia coli DmsD interacts with the twin-arginine leader sequence of the catalytic sub-unit (DmsA) of DMSO reductase. DmsD was purified as a mixture of a number of different folding forms including: dimer (A); monomer (B); a minor thiol oxidized form; a heterogeneously folded or multi-conformational monomer form which displayed a ladder of bands on native-PAGE (D); and proteolytically degraded and aggregated forms. Polyacrylamide gel electrophoresis (PAGE), under denaturing and non-denaturing conditions, was used to examine the folding and stability of DmsD. Additionally, the biophysical methods of dynamic light scattering, circular dichroism, fluorescence, and mass spectroscopy were also used. Form D could be converted to form B by treatment with 4M urea, which is the concentration at which form B begins to denature. Forms A/B could be converted to D by incubation at pH 5.0. Forms A/B and D all had twin-arginine leader binding activity.
