Fr e superbright generator

C. thermophilum Vps29 is 201 amino acids in length and
thrombin cleavage leaves an additional two non-native N-terminal residues
(GlySer). Crystals of Vps29 were produced by hanging drop vapour diffusion using
protein at a concentration of 15 mg/ml. Crystals in spacegroup P2₁grew from a reservoir solution of 8% PEG20000, 8% PEG550MME, 0.2 M calcium
acetate and 0.1 M Tris (pH 8.0), and were cryoprotected in 25% glycerol.
Crystals were screened at the UQ ROCX diffraction facility on a Rigaku FR-E
Superbright generator with Osmic Vari-Max HF optics and Rigaku Saturn 944 CCD
detector. Data for structure determination was collected at the Australian
Synchrotron MX1 Beamline. Data was integrated with iMOSFLM30 (link) and scaled with SCALA31 (link). The structure was solved by molecular replacement with
PHASER32 (link) using the human Vps29
protein18 (link) as an input model. The
resulting models were rebuilt with COOT33 (link)
and refined with PHENIX34 (link).
Crystallographic data and structure statistics are provided in Extended Data Table 2.

Ldt_Mt2-apo, co-crystals as well as crystals soaked with carbapenem drugs were cryo-protected in 30% glycerol, 20% 5000 MME and 120 mM ammonium sulfate buffer and flash cooled in liquid nitrogen. For Ldt_Mt2-apo and co-crystals with faropenem and doripenem, X-ray diffraction data were collected at a wavelength of 1.54 Å using an in-house CuKα X-ray source (Rigaku FR-E+ SuperBright generator with a Saturn 944+ CCD Detector; Rigaku). for Ldt_Mt2 apo and crystals soaked with evolved the carbapenems T206, T208 and T210, diffraction data were collected at 100K at a wavelength of 0.98 Å on beamline 19-ID at the Advanced Photon Source (Argonne National Laboratory) and the diffraction data were recorded on an ADSC Quantum 315r CCD detector, and processed with HKL3000 ^{38 (link)}.
Ldt_Mt1-apo and co-crystal with faropenem were cryoprotected with 30% PEG8000, 10% PEG6000 and 100 mM Bicine pH 9.0 buffer and flash cooled in liquid nitrogen. The X-ray diffraction data were collected and analyzed as described below.