Heterologous overexpression of a monotopic glucosyltransferase (MGS) induces fatty acid remodeling in Escherichia coli membranes

• Published in: Biochimica et biophysica acta 2014-07, Vol.1838 (7), p.1862-1870
• Main Authors: Ariöz, Candan, Götzke, Hansjörg, Lindholm, Ljubica, Eriksson, Jonny, Edwards, Katarina, Daley, Daniel O., Barth, Andreas, Wieslander, Åke
• Format: Article
• Language: English
• Subjects: Acholeplasma laidlawii - enzymology; Acholeplasma laidlawii - genetics; Acholeplasma laidlawii - metabolism; Biochemistry; biokemi; Cell Membrane - enzymology; Cell Membrane - metabolism; Cyclopropanation; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli - metabolism; Fatty acid modeling; Fatty Acids - metabolism; Glucosyltransferases - metabolism; Intracellular membrane; intracellular membranes; Lipid–protein interaction; Lipid–protein interactions; Membrane fluidity; Membrane Lipids - metabolism; Membrane Proteins - metabolism; Methyltransferases - metabolism; Models, Molecular; Monotopic membrane protein; monotopic membrane proteins; Phosphatidylethanolamines - metabolism; Protein Structure, Secondary
• ISSN: 0005-2736; 0006-3002; 1879-2642; 1879-2642
• DOI: 10.1016/j.bbamem.2014.04.001

The membrane protein monoglucosyldiacylglycerol synthase (MGS) from Acholeplasma laidlawii is responsible for the creation of intracellular membranes when overexpressed in Escherichia coli (E. coli). The present study investigates time dependent changes in composition and properties of E. coli membranes during 22h of MGS induction. The lipid/protein ratio increased by 38% in MGS-expressing cells compared to control cells. Time-dependent screening of lipids during this period indicated differences in fatty acid modeling. (1) Unsaturation levels remained constant for MGS cells (~62%) but significantly decreased in control cells (from 61% to 36%). (2) Cyclopropanated fatty acid content was lower in MGS producing cells while control cells had an increased cyclopropanation activity. Among all lipids, phosphatidylethanolamine (PE) was detected to be the most affected species in terms of cyclopropanation. Higher levels of unsaturation, lowered cyclopropanation levels and decreased transcription of the gene for cyclopropane fatty acid synthase (CFA) all indicate the tendency of the MGS protein to force E. coli membranes to alter its usual fatty acid composition. [Display omitted] •Overexpression of MGS in Escherichia coli membranes changes fatty acid composition.•Cyclopropanation is substantially decreased by MGS expression.•cfa gene transcription is lowered in MGS expressing cells.•BL21 AI and BL21 AI-MGS cells both maintain a liquid crystalline phase.