title: Cold-adapted protease enables quantitation of surface proteins in the absence of membrane trafficking.
abstract: We report here an improved method for analyzing protein surface expression utilizing a cold-adapted trypsin. Preservation of activity of the enzyme at 0-4°C permits modification of the protease method of surface analysis to temperatures at which trafficking of mammalian plasmalemmal proteins is blocked. This is an important advantage over established trypsin-cleavage protocols. Moreover, the method is less time-consuming than surface biotinylation.
authors.affiliation: Laboratory of Neurobiology, University of Helsinki, Helsinki, Finland.
authors.forename: Faraz
authors.initials: F
authors.lastname: Ahmad
authors.forename: Sarah Kate
authors.initials: SK
authors.lastname: Coleman
authors.forename: Kai
authors.initials: K
authors.lastname: Kaila
authors.forename: Peter
authors.initials: P
authors.lastname: Blaesse
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry* / Q000201:enzymology; D003080:Cold Temperature; D006624:Hippocampus / Q000737:chemistry / Q000378:metabolism; D015151:Immunoblotting; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D017470:Receptors, Glutamate / Q000032:analysis / Q000378:metabolism; D013499:Surface Properties; D027981:Symporters / Q000032:analysis / Q000378:metabolism; D014357:Trypsin / Q000737:chemistry* / Q000378:metabolism; D000096922:K Cl- Cotransporters
chemical_list: D008565:Membrane Proteins; D017470:Receptors, Glutamate; D027981:Symporters; D014357:Trypsin; D000096922:K Cl- Cotransporters