Automated Subunit Counting of Membrane Proteins 1 AUTOMATING SINGLE SUBUNIT COUNTING OF MEMBRANE PROTEINS IN MAMMALIAN CELLS*
نویسندگان
چکیده
Background: Although powerful, single subunit counting is time consuming, prone to user-bias and largely restricted to Xenopus expression. Results: PIF is an automated analysis program that identifies subunit stoichiometry of any fluorescently-tagged membrane protein from TIRF-recordings. Conclusion: PIF is accurate to more than 90% even in noisy data typical for mammalian expression system. Significance: PIF approach is generalizable to any membrane protein and TIRF-microscope.
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Automating single subunit counting of membrane proteins in mammalian cells.
Elucidating subunit stoichiometry of neurotransmitter receptors is preferably carried out in a mammalian expression system where the rules of native protein assembly are strictly obeyed. Although successful in Xenopus oocytes, single subunit counting, manually counting photobleaching steps of GFP-tagged subunits, has been hindered in mammalian cells by high background fluorescence, poor control...
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