SAXS TOWARD STRUCTURE DETERMINATION OF ACETOACETATE DECAROXYLASE

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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. Acetoacetate Deacrboxylase is the first enzyme to be studied mechanistically, and to be used in a biotech application for producing acetone. An EM map obtained to 9.8 angstrom resolution shows a highly unusual assembly for this dodecameric enzyme- a hollow sphere with D6 symmetry, 100 angstroms in diameter made up of all beta sheet. The proposed SAXS experiment will allow us to modulate the contrast function for EM and to measure the precise shape and volume of one sample in order to re-scale our EM data. The long range goal is to use the EM phases to phase our high resolutuion (1.8 angstrom) X-ray data. As the phasing process is held up until SAXS can be done, rapid access is important.