An experimental approach to detect the path a substrate takes through a complex membrane protein is described with emphasis on technical approach and theoretical considerations. The protocols for bacterial culture preparation, membrane protein purification, fluorescent assay standardization, data collection, and data analysis are provided. Useful software tools are recommended.
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I am thankful to Dr. Etsuko Sugawara for suggestions and Dr. Yumiko Takatsuka and Dr. Rajeev Misra for materials. This work was carried out in the laboratory of Dr. Hiroshi Nikaido at University of California, supported by AI-09644 grant from U.S. Public Health Service.
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