Purification and characterization of the bacterial UDP-GlcNAc:undecaprenyl-phosphate GlcNAc-1-phosphate transferase WecA.

Journal of Bacteriology
Bayan Al-DabbaghAhmed Bouhss

Abstract

To date, the structural and functional characterization of proteins belonging to the polyprenyl-phosphate N-acetylhexosamine-1-phosphate transferase superfamily has been relentlessly held back by problems encountered with their overexpression and purification. In the present work and for the first time, the integral membrane protein WecA that catalyzes the transfer of the GlcNAc-1-phosphate moiety from UDP-GlcNAc onto the carrier lipid undecaprenyl phosphate, yielding undecaprenyl-pyrophosphoryl-GlcNAc, the lipid intermediate involved in the synthesis of various bacterial cell envelope components, was overproduced and purified to near homogeneity in milligram quantities. An enzymatic assay was developed, and the kinetic parameters of WecA as well as the effects of pH, salts, cations, detergents, and temperature on the enzyme activity were determined. A minimal length of 35 carbons was required for the lipid substrate, and tunicamycin was shown to inhibit the enzyme at submicromolar concentrations.

References

May 1, 1995·Trends in Microbiology·C Whitfield
Jan 8, 1999·Biochimica Et Biophysica Acta·P Burda, M Aebi
Jun 12, 2002·Journal of Bacteriology·Christopher K RaymondMaynard V Olson
Apr 18, 2003·Frontiers in Bioscience : a Journal and Virtual Library·Miguel A Valvano
Jun 26, 2003·Protein Science : a Publication of the Protein Society·Jing Liu, Arcady Mushegian
May 13, 2004·The Journal of Biological Chemistry·Meriem El GhachiDominique Mengin-Lecreulx
Apr 22, 2005·Glycobiology·Neil P Price, Frank A Momany
Sep 6, 2006·Journal of Bacteriology·Katarína MikusováPatrick J Brennan
Dec 18, 2007·FEMS Microbiology Reviews·Ahmed BouhssDominique Mengin-Lecreulx
Feb 22, 2008·Current Topics in Medicinal Chemistry·Hongjie GuoPeng George Wang

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Citations

Nov 17, 2011·Journal of Medicinal Chemistry·Tetsuya TaninoAkira Matsuda
Apr 27, 2010·Nature Chemical Biology·Robert WoodwardPeng George Wang
Dec 3, 2014·International Journal of Molecular Sciences·Susana MerinoJuan M Tomás
Mar 25, 2010·Biochemical and Biophysical Research Communications·Lei LiPeng George Wang
Dec 15, 2015·Chemistry : a European Journal·Marthe T C WalvoortBarbara Imperiali
Aug 30, 2016·Antibiotics·Yao Liu, Eefjan Breukink
Oct 22, 2016·European Journal of Medicinal Chemistry·Michaël BoscoChristine Gravier-Pelletier
Jun 21, 2017·Proceedings of the National Academy of Sciences of the United States of America·Debasis DasBarbara Imperiali
Sep 7, 2017·Antimicrobial Agents and Chemotherapy·Stanislav HuszárKatarína Mikušová
Sep 4, 2014·Microbiology and Molecular Biology Reviews : MMBR·Hanne L P Tytgat, Sarah Lebeer
Oct 30, 2016·Applied and Environmental Microbiology·Tuan Ngoc NguyenShir-Ly Huang
Sep 1, 2019·The Journal of Antibiotics·Yoshimasa IshizakiMasayuki Igarashi
Oct 1, 2019·Nucleosides, Nucleotides & Nucleic Acids·Kazuki YamamotoSatoshi Ichikawa
Jun 23, 2020·Annual Review of Biochemistry·Christopher A CaffaletteJochen Zimmer
May 27, 2016·The Journal of Biological Chemistry·Yao LiuMaarten R Egmond
Feb 27, 2018·PLoS Pathogens·Paul A BeareRobert A Heinzen
Jun 27, 2019·The Journal of Antibiotics·Kazuki Yamamoto, Satoshi Ichikawa

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