EXPRESSION AND CRYSTALLIZATION OF MYCOBACTERIAL GALACTOFURANOSYL TRANSFERASE GLFT1

KYSEĽ, Peter, Gabriel DEMO, Stanislav HUSZÁR, Katarina MIKUŠOVÁ and Michaela WIMMEROVÁ. EXPRESSION AND CRYSTALLIZATION OF MYCOBACTERIAL GALACTOFURANOSYL TRANSFERASE GLFT1. In XVII. setkání biochemiků a molekulárních biologů. 2015. ISBN 978-80-210-8015-7.

Basic information

• Original name: EXPRESSION AND CRYSTALLIZATION OF MYCOBACTERIAL GALACTOFURANOSYL TRANSFERASE GLFT1
• Authors: KYSEĽ, Peter (703 Slovakia, belonging to the institution), Gabriel DEMO (703 Slovakia, belonging to the institution), Stanislav HUSZÁR (703 Slovakia), Katarina MIKUŠOVÁ (703 Slovakia) and Michaela WIMMEROVÁ (203 Czech Republic).
• Edition: XVII. setkání biochemiků a molekulárních biologů, 2015.

Other information

• Original language: English
• Type of outcome: Conference abstract
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: Czech Republic
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• RIV identification code: RIV/00216224:14310/15:00085288
• Organization unit: Faculty of Science
• ISBN: 978-80-210-8015-7
• Keywords in English: mycobacterium; glycosyltransferase; purification; detergent;crystallography
• Tags: AKR

Abstract

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.

Links

• ED1.1.00/02.0068, research and development project: Name: CEITEC - central european institute of technology