TRANSIENT STRUCTURAL STATES OF GROEL

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. ABSTRACT A collaborative project (Art Horwich & George Farr, Yale) to investigate transient structural states of the chaperonin, GroEL, has commenced. This project will make use of the pre-mix/spray and flash-photolysis modes of time-resolved cryo-EM. We have described our implementation of flash-photolysis in previous reports. One problem that remains not fully resolved is the degree to which the temperature of the reaction solution rises upon exposure to a UV light flash which initiates photolysis of caged reactants. Currently, to minimize the temperature rise we use 50-um thick copper grids (custom made by Ladd Research) that are coated with aluminum and then covered with carbon nets to minimize absorption of photons by the carbon.