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EXPRESSION AND CRYSTALLIZATION OF MYCOBACTERIAL GALACTOFURANOSYL TRANSFERASE GLFT1

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KYSEĽ Peter DEMO Gabriel HUSZÁR Stanislav MIKUŠOVÁ Katarina WIMMEROVÁ Michaela

Druh Konferenční abstrakty
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Popis A number of glycosyltransferases is involved in the mycobacterial cell wall biosynthesis. The membrane associated galactofuranosyltransferase GlfT1 adds the first two galactofuranosyl residues of the galactan component of the arabinogalactan. Yet the esentiality of the GlfT1 for the proliferation of persistent mycobacterial pathogens provided a good reason to compose and optimise a combination of different biochemical approaches for purification and crystallization of this enzyme. We developed a multistep purification procedure to counter the pitfalls of proteolysis, chaperone contamination, precipitation and polydispersity. Two different conditions were identified to faciliate the crystallization of the fusion protein. X-ray diffraction data to 2.1 A were obtained. Solving of GlfT1 3D structure will eventually deliver structural insights into the first steps of galactan biosynthesis.
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