HIGH RESOLUTION DATA SET OF 2 FORMS OF VANCOMYCIN RESISTANCE ACCOCIATED

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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. 1) Vancomycin resistance accociated Regulator (VraR) is involved in mounting a response to antibiotices that target cell wall peptidoglycan biosynthesis such as vancomycin. This enzyme is not present in mammals and thus becomes a good target for a drug to target vancomycin resistant bacteria. (2 MAD data sets + 2 high resolution) (1 MAD set and high res is absolutely needed now, the highest priority) The other MAD set and high res is the second thing that we need from this trip. Also, 2 high resolution sets of a different space group would be collected last on the trip. Only if there is extra time, this is not a primary data set. All of these crystals have been frozen and pre-screened on our rotating anode --------------------------------------------------------------- 2) E. coli serine protease (ECSP) cleaves several significant proteins. We have previously solved the structure and would now like to look at consensus peptides bound to the active site. 2 data sets - The 2.5A was low exposure just to check the crystal quality of pre-frzen crystals. Crystals are ordered to better than 2A. All of these crystals have been frozen and pre-screened on our rotating anode. These will diffract to higher resolution at CHESS. The native protein diffracts much better, better than 2A. We are trying to get some additional similar standard peptides into the protein. If these work before the trip, we will pre-freeze and pre-screen them and bring them along to collect. -------------------------------------------------------------- 3) (S)-3-O-Geranylgeranlyglyceryl Phosphate Synthase (GGGPS) is isolated from Archaea and is involved in synthesis of compounds unique to Archaea cell walls. We would like to look at the mechanism of this unique type of enzymes. 3 data sets - they have been frozen and pre-screened on our rotating anode and stored for the trip. There are a few other similar mutants being crystallized. If we can get crystals, pre-freeze and pre-test them, we will bring them along to collect, if there is time. ------------------------------------------------------------------ Every crystal on our list has been pre-frozen and pre-screened on our rotating anode. We will bring some room temperature plates, but purely as a backup. The MAD data on VraR is our priority for this trip.