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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Membrane / chemistry*
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Cell Membrane / enzymology
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Cold Temperature*
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Hippocampus / chemistry
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Hippocampus / metabolism
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Immunoblotting
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K Cl- Cotransporters
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Membrane Proteins / analysis*
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Membrane Proteins / chemistry
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Membrane Proteins / metabolism
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Rats
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Receptors, Glutamate / analysis
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Receptors, Glutamate / metabolism
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Surface Properties
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Symporters / analysis
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Symporters / metabolism
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Trypsin / chemistry*
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Trypsin / metabolism
Substances
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Membrane Proteins
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Receptors, Glutamate
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Symporters
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Trypsin