Membrane Interaction of the Glycosyltransferase MurG: a Special Role for Cardiolipin

MurG is a peripheral membrane protein that is one of the key enzymes in peptidoglycan biosynthesis. The crystal structure of Escherichia coli MurG (S. Ha, D. Walker, Y. Shi, and S. Walker, Protein Sci. 9:1045-1052, 2000) contains a hydrophobic patch surrounded by basic residues that may represent a...

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Hlavní autoři: van den Brink-van der Laan, Els, Boots, Jan-Willem P., Spelbrink, Robin E. J., Kool, Gerda M., Breukink, Eefjan, Killian, J. Antoinette, de Kruijff, Ben
Médium: Článek
Jazyk:en
Vydáno: American Society for Microbiology 2003
Témata:
Enzymes and Proteins
On-line přístup:https://ncbi.nlm.nih.gov/pmc/articles/PMC161595/
https://ncbi.nlm.nih.gov/pubmed/12813070
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1128/JB.185.13.3773-3779.2003
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spelling pubmed-1615952003-07-29 Membrane Interaction of the Glycosyltransferase MurG: a Special Role for Cardiolipin van den Brink-van der Laan, Els Boots, Jan-Willem P. Spelbrink, Robin E. J. Kool, Gerda M. Breukink, Eefjan Killian, J. Antoinette de Kruijff, Ben J Bacteriol Enzymes and Proteins MurG is a peripheral membrane protein that is one of the key enzymes in peptidoglycan biosynthesis. The crystal structure of Escherichia coli MurG (S. Ha, D. Walker, Y. Shi, and S. Walker, Protein Sci. 9:1045-1052, 2000) contains a hydrophobic patch surrounded by basic residues that may represent a membrane association site. To allow investigation of the membrane interaction of MurG on a molecular level, we expressed and purified MurG from E. coli in the absence of detergent. Surprisingly, we found that lipid vesicles copurify with MurG. Freeze fracture electron microscopy of whole cells and lysates suggested that these vesicles are derived from vesicular intracellular membranes that are formed during overexpression. This is the first study which shows that overexpression of a peripheral membrane protein results in formation of additional membranes within the cell. The cardiolipin content of cells overexpressing MurG was increased from 1 ± 1 to 7 ± 1 mol% compared to nonoverexpressing cells. The lipids that copurify with MurG were even further enriched in cardiolipin (13 ± 4 mol%). MurG activity measurements of lipid I, its natural substrate, incorporated in pure lipid vesicles showed that the MurG activity is higher for vesicles containing cardiolipin than for vesicles with phosphatidylglycerol. These findings support the suggestion that MurG interacts with phospholipids of the bacterial membrane. In addition, the results show a special role for cardiolipin in the MurG-membrane interaction. American Society for Microbiology 2003-07 /pmc/articles/PMC161595/ /pubmed/12813070 http://dx.doi.org/10.1128/JB.185.13.3773-3779.2003 Text en Copyright © 2003, American Society for Microbiology
institution US National Library of Medicine
collection PubMed Central
language en
format Article
topic Enzymes and Proteins
spellingShingle Enzymes and Proteins
van den Brink-van der Laan, Els
Boots, Jan-Willem P.
Spelbrink, Robin E. J.
Kool, Gerda M.
Breukink, Eefjan
Killian, J. Antoinette
de Kruijff, Ben
Membrane Interaction of the Glycosyltransferase MurG: a Special Role for Cardiolipin
description MurG is a peripheral membrane protein that is one of the key enzymes in peptidoglycan biosynthesis. The crystal structure of Escherichia coli MurG (S. Ha, D. Walker, Y. Shi, and S. Walker, Protein Sci. 9:1045-1052, 2000) contains a hydrophobic patch surrounded by basic residues that may represent a membrane association site. To allow investigation of the membrane interaction of MurG on a molecular level, we expressed and purified MurG from E. coli in the absence of detergent. Surprisingly, we found that lipid vesicles copurify with MurG. Freeze fracture electron microscopy of whole cells and lysates suggested that these vesicles are derived from vesicular intracellular membranes that are formed during overexpression. This is the first study which shows that overexpression of a peripheral membrane protein results in formation of additional membranes within the cell. The cardiolipin content of cells overexpressing MurG was increased from 1 ± 1 to 7 ± 1 mol% compared to nonoverexpressing cells. The lipids that copurify with MurG were even further enriched in cardiolipin (13 ± 4 mol%). MurG activity measurements of lipid I, its natural substrate, incorporated in pure lipid vesicles showed that the MurG activity is higher for vesicles containing cardiolipin than for vesicles with phosphatidylglycerol. These findings support the suggestion that MurG interacts with phospholipids of the bacterial membrane. In addition, the results show a special role for cardiolipin in the MurG-membrane interaction.
author van den Brink-van der Laan, Els
Boots, Jan-Willem P.
Spelbrink, Robin E. J.
Kool, Gerda M.
Breukink, Eefjan
Killian, J. Antoinette
de Kruijff, Ben
author_facet van den Brink-van der Laan, Els
Boots, Jan-Willem P.
Spelbrink, Robin E. J.
Kool, Gerda M.
Breukink, Eefjan
Killian, J. Antoinette
de Kruijff, Ben
author_sort van den Brink-van der Laan, Els
title Membrane Interaction of the Glycosyltransferase MurG: a Special Role for Cardiolipin
title_short Membrane Interaction of the Glycosyltransferase MurG: a Special Role for Cardiolipin
title_full Membrane Interaction of the Glycosyltransferase MurG: a Special Role for Cardiolipin
title_fullStr Membrane Interaction of the Glycosyltransferase MurG: a Special Role for Cardiolipin
title_full_unstemmed Membrane Interaction of the Glycosyltransferase MurG: a Special Role for Cardiolipin
title_sort membrane interaction of the glycosyltransferase murg: a special role for cardiolipin
publisher American Society for Microbiology
publisher_facet American Society for Microbiology
publishDate 2003
url https://ncbi.nlm.nih.gov/pmc/articles/PMC161595/
https://ncbi.nlm.nih.gov/pubmed/12813070
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1128/JB.185.13.3773-3779.2003
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