O. Till, M. Schmidt, W. Linss
Mar 1, 2007
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0
Influential Citations
2
Citations
Quality indicators
Journal
ELECTROPHORESIS
Abstract
The applicability of the membrane‐impermeant protein cross‐linker bis(sulfosuccinimidyl) suberate (BS3) to the determination of membrane sidedness of proteins was tested in 3T3‐L1 cells and in erythrocytes. Binding of BS3 to proteins was apparent in electrophoresis. In three proteins of 3T3‐L1 cells, protein kinase‐Cϵ, protein kinase‐Cζ, and glyceraldehyde‐3‐phosphate dehydrogenase, BS3 action was detectable in SDS‐PAGE with immunoblotting. This enabled confirmation of the well‐known intracellular localization of these proteins. In cathepsin E of erythrocytes, a mobility increase in nondenaturing PAGE was the most prominent effect of BS3 treatment. A mechanism for the increase in mobility due to BS3 binding is suggested. Cathepsin E was found to be located at the intracellular side of the membrane, in accordance with existing evidence.