Cold-adapted protease enables quantitation of surface proteins in the absence of membrane trafficking. (April 2011)

• Record Type: Journal Article
• Authors: Ahmad, Faraz; Coleman, Sarah Kate; Kaila, Kai; Blaesse, Peter
• Is Part Of: Biotechniques. Volume 50:Number 4(2011)
• Page Start: 255
• Page End: 257
• Publication Date: 2011-04
• Subjects: membrane protein -- surface expression -- trafficking -- trypsin -- biotinylation -- KCC2 -- GluA4; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000113651
• Languages: English
• ISSNs: 0736-6205

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.