EXPRESSION AND CRYSTALLIZATION OF MYCOBACTERIAL
GALACTOFURANOSYL TRANSFERASE GLFT1

a 2015

EXPRESSION AND CRYSTALLIZATION OF MYCOBACTERIAL GALACTOFURANOSYL TRANSFERASE GLFT1

KYSEĽ, Peter, Gabriel DEMO, Stanislav HUSZÁR, Katarina MIKUŠOVÁ, Michaela WIMMEROVÁ et. al.

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

Language

English

Type of outcome

Konferenční abstrakt

Field of Study

10600 1.6 Biological sciences

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

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

Abstract

V originále

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

Citovat

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.

@proceedings{1321653,
   author = {Kyseľ, Peter and Demo, Gabriel and Huszár, Stanislav and Mikušová, Katarina and Wimmerová, Michaela},
   booktitle = {XVII. setkání biochemiků a molekulárních biologů},
   keywords = {mycobacterium; glycosyltransferase; purification; detergent;crystallography},
   language = {eng},
   isbn = {978-80-210-8015-7},
   title = {EXPRESSION AND CRYSTALLIZATION OF MYCOBACTERIAL GALACTOFURANOSYL TRANSFERASE GLFT1},
   url = {http://orion.chemi.muni.cz/setkani/Setkani17/index.htm},
   year = {2015}
}

TY  - CONF
ID  - 1321653
AU  - Kyseľ, Peter - Demo, Gabriel - Huszár, Stanislav - Mikušová, Katarina - Wimmerová, Michaela
PY  - 2015
TI  - EXPRESSION AND CRYSTALLIZATION OF MYCOBACTERIAL GALACTOFURANOSYL TRANSFERASE GLFT1
SN  - 9788021080157
KW  - mycobacterium
KW  - glycosyltransferase
KW  - purification
KW  - detergent;crystallography
UR  - http://orion.chemi.muni.cz/setkani/Setkani17/index.htm
L2  - http://orion.chemi.muni.cz/setkani/Setkani17/index.htm
N2  - 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.
ER  -

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

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