Glycerol-3-phosphate cytidylyltransferase from ππ΅π’π±π©πΊππ°π€π°π€π€πΆπ΄ π’πΆπ³π¦πΆπ΄ complexed with CTP (TarDββ-CTP) was crystallized by the hanging drop-vapor diffusion method at 22Β°C. Determination of crystallization condition included examination of the amount of precipitant, investigation of the effects of small molecules, and alteration of the rate of diffusion. With these three optimization steps, crystals suitable for x-ray diffraction study were produced. During data processing, TarDββ-CTP was determined to belong to the space group P3β21, with unit-cell dimensions a=b=92.2 and c=156.1Γ . The crystal structure of TarDββ-CTP was solved to 3.0Γ by molecular replacement, using TagD from ππ’π€πͺπππΆπ΄ π΄πΆπ£π΅πͺππͺπ΄ as a search model. Unlike the search model, TarDββ appears as a tetramer in the asymmetric unit. This result also confirms the gel-filtration and ultracentrifugation studies that were done previously. Although TarDββ crystals were grown in the presence of CTP, the crystal structure does not reveal convincing data for the location and position of this co-factor. However, the data suggests a possible location for CTP in one of the four subunits in an orientation that differs from that of TagD_Bβ. Unfortunately, the resolution of this data set at 3.0Γ is not high enough to corroborate this finding. / Thesis / Master of Science (MS)
| Identifer | oai:union.ndltd.org:mcmaster.ca/oai:macsphere.mcmaster.ca:11375/22976 |
| Date | January 2002 |
| Creators | Yim, Veronica |
| Contributors | Berghuis, A. M., Biochemistry |
| Source Sets | McMaster University |
| Language | English |
| Detected Language | English |
| Type | Thesis |
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