Publication details

EXPRESSION AND CRYSTALLIZATION OF MYCOBACTERIAL GALACTOFURANOSYL TRANSFERASE GLFT1

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Authors

KYSEĽ Peter DEMO Gabriel HUSZÁR Stanislav MIKUŠOVÁ Katarina WIMMEROVÁ Michaela

Year of publication 2015
Type Conference abstract
MU Faculty or unit

Faculty of Science

Citation
Description 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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