Proteinase K + surface localization of proteins? - (May/15/2007 )
Anyone that knows a bit about this topic? I would like to use Proteinase K in an assay to help me understand better possible surface localization of a bacterial protein I am after, so I was wondering how can this assay help me answer this? I've read most of literature about Proteinase K and I still don't get it. If my protein seems resistant to proteolytic cleavage by Proteinase K, does that mean it is located in the membrane? So if it is surface located, it should be susceptible to this hydrolysis and no protein band should appear in the Western Blot?
well i would try to prepare membrane proteins extract. you should be able to know if the protein seems to be in the memebrane of the cell. If your protein is in membrane there should be a part exposed to sirface which should be cleavable by the proteinase K.
May you try a flagged protein and do immuno fluo against the tag to see if it's near the surface, and would disappear after treatment by prot K ?