Journal
JOURNAL OF PHYSICAL CHEMISTRY B
Volume 118, Issue 11, Pages 2949-2956Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jp412631d
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Funding
- Department of Science and Technology, India [IR/S1/CU 02/2009]
- Council for Scientific and Industrial Research (CSIR)
- J. C. Bose Fellowship
- CSIR
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Dynamics of the exofacial thiols (i.e., cell surface thiol containing membrane proteins) of a live Chinese hamster ovary (CHO) cell is probed by time-resolved confocal microscopy. For this purpose, a fluorescent probe, 7-(diethylamino)-3-(4-maleimidophenyl)-4-methylcoumarin (CPM) is covalently attached to the exofacial thiols. The emission maximum of CPM bound exofacial thiols indicates a highly exposed and polar environment. Using CPM, we studied solvation dynamics, for the first time, at the membrane of a live cell. The thiol containing membrane proteins shows ultraslow response with average solvation time, = 475 ps. CPM labeled exofacial thiols also show spontaneous, intermittent oscillation in fluorescence intensity with a period of 0.5-1.0 s. This is ascribed to reversible, intermittent changes in the structure and conformation of the membrane proteins.
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