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

Laan, Els van den Brink-van der; Boots, Jan-Willem P.; Spelbrink, Robin E.J.; Kool, Gerda M.; Breukink, Eefjan; Killian, J. Antoinette; Kruijff, Ben de

doi:10.1128/jb.185.13.3773-3779.2003

Cited by 73 publications

(72 citation statements)

References 33 publications

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“…In general, the membrane proteins known to be CL-dependent are channels or transporters that use multiple membrane domains to constitute a pore within the bilayer. An interesting exception is MurG, an E. coli glycosyltransferase involved in peptidoglycan biosynthesis (14). Overexpression of this peripheral membrane protein results in elevated CL content in cell membranes and association of the protein with unusual intracellular vesicles.…”

Section: Discussionmentioning

confidence: 99%

“…The activity of many integral membrane enzymes is dependent on, or influenced by, a specific type of phospholipid, and there are now many cases known in which the activating lipid is CL (13)(14)(15)(16)(17)(18). Mitochondrial enzymes involved in oxidative phosphorylation comprise a large fraction of this group, and their ability to form large functional complexes is CL-dependent.…”

Section: Hyaluronan Synthase (Has)mentioning

confidence: 99%

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Phospholipid Dependence and Liposome Reconstitution of Purified Hyaluronan Synthase

Weigel

Kyossev

Torres

2006

Journal of Biological Chemistry

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Previous radiation inactivation and enzyme characterization studies demonstrated that the Streptococcus equisimilis hyaluronan synthase (seHAS) is phospholipid-dependent and that cardiolipin (CL) is the best phospholipid for enzyme activation. Here we investigated the ability of seHAS, purified in the absence of added lipid, to be activated by synthetic phosphatidic acid (PA), phosphatidylserine, or CL lipids containing fatty acyl chains of different length or different numbers of double bonds. The most effective lipid was tetraoleoyl CL (TO-CL), whereas tetramyristoyl CL (TM-CL) was ineffective. None of the phosphatidylserine species tested gave significant activation. PAs containing C10 to C18 saturated acyl chains were not effective activators, and neither were oleoyl lyso PA, dilinoleoyl PA, or PA containing one oleoyl chain and either a palmitoyl or stearoyl chain. In contrast, dioleoyl PA stimulated seHAS ϳ10-fold, to ϳ20% of the activity observed with TO-CL. The tested acidic lipids such as PA and CL activated the enzyme most efficiently if they contained only oleic acid. Mixing experiments showed that the enzyme interacts preferentially with TO-CL in the presence of TM-CL. Similarly, seHAS incorporated into phosphotidylcholine-based liposomes showed increasing activity with increasing TO-CL, but not TM-CL, content. Inactivation of membrane-bound seHAS by solubilization with Nonidet P-40 was prevented by TO-CL, but not TM-CL. The pH dependence of seHAS in the presence of synthetic or naturally occurring CLs showed the same pattern of lipid preference between pH 6 and 10.5. Unexpectedly, HAS showed lipid-independent activity at pH 11.5. The results suggest that Class I HAS enzymes are lipid-dependent and that assembly of active seHAS-lipid complexes has high specificity for the phospholipid head group and the nature of the fatty acyl chains.

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Section: Discussionmentioning

confidence: 99%

Section: Hyaluronan Synthase (Has)mentioning

confidence: 99%

Phospholipid Dependence and Liposome Reconstitution of Purified Hyaluronan Synthase

Weigel

Kyossev

Torres

2006

Journal of Biological Chemistry

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“…Specialization is not associated with insertions at cls (including strains adapted to mixed sugars, P ϭ 0.08 by a one-tailed Fisher's exact test) or with type II deletions (including strains adapted to mixed sugars, P ϭ 0.14 by a one-tailed Fisher's exact test). Insertional inactivation of cls, which encodes cardiolipin synthase (48), affects many membrane functions (49)(50)(51)(52), any one of which might be a target of selection. Motility is pointless in the well-stirred environment of a chemostat.…”

Section: Discussionmentioning

confidence: 99%

Evolutionary genomics of ecological specialization

Zhong

Khodursky

Dykhuizen

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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We used a combination of genomic techniques to monitor chromosomal evolution across hundreds of generations as Escherichia coli adapted to growth-limiting concentrations of either lactulose, methyl-galactoside, or a 72:28 mixture of the two. DNA microarrays identified 8 unique duplications and 16 unique deletions among 42 evolvants from 23 chemostat experiments. Each mutation was confirmed by sequencing PCR-amplified flanking genomic DNA and, except for one deletion, an insertion sequence was found at the break point. vPCR of insertion sequences identified these same mutations and 16 additional insertions (all confirmed by sequencing). The pattern of genomic evolution is highly reproducible. Statistical analyses show that duplications at lac and mutations in mgl are adaptations specific to lactulose and to methyl-galactoside, respectively. Adaptation to mixed sugars is characterized by similar mutations, but lac duplications and mgl mutations usually arise in different backgrounds, producing ecological specialists for each sugar. This suggests that an antagonistic pleiotropic tradeoff between duplications at lac and mutations in mgl retards the evolution of generalists. Other mutations that repeatedly appear in replicate experiments are adaptations to the chemostat environment and are not specific to one or the other sugar.W e have taken advantage of the repeatability of laboratory adaptation to investigate the roles of insertion sequences (IS) in the evolution of resource specialization.The repeatability of laboratory adaptation is well documented (1-11) and is perhaps attributable to using large populations with simple ecologies. Large populations reduce the role of chance in evolution: random genetic drift is minimized, whereas mutation, always erratic in small populations, produces numerous allelic variants with each generation. In simple constant environments, selection becomes focused with great intensity on a few key genes. This makes experimental evolution far more reproducible than typically envisioned for small populations inhabiting complex natural environments (12). Reproducibility makes possible rigorous tests of adaptive hypotheses. For example, Cooper et al. (13) used DNA expression macroarrays to identify parallel changes in the expression profiles of two evolvants. Guided by the observation that many of the genes with changed expression belong to the ppGpp and CRP regulons, they identified a mutation in spoT in one population that, when reintroduced into the ancestral genetic background, increased fitness as well as reproducing many of the changes in expression.IS elements are small (1-to 2-kb) segments of DNA capable of transposing within and between prokaryotic replicons (14). A major source of insertional mutations and chromosomal rearrangements (15), their evolutionary significance is a subject of perennial interest (16)(17)(18)(19)(20). Patterns of sequence polymorphism among natural isolates of Escherichia coli suggest a brisk turnover of elements (21), with both the numbers and location...

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“…Interface-interacting Proteins-Several monotopic membrane proteins also induce membrane formation, like the E. coli glycerol-3-phosphate acyltransferase PlsB (8), the peptidoglycan precursor glycosyltransferase MurG with an established structure (39,55), and the two heterologous (A. laidlawii) lipid glycosyltransferases alMGS and alDGS in this work (cf. results above).…”

Section: Formation Of Intracellular Membranesmentioning

confidence: 99%

Massive Formation of Intracellular Membrane Vesicles in Escherichia coli by a Monotopic Membrane-bound Lipid Glycosyltransferase

Eriksson

Wessman

et al. 2009

Journal of Biological Chemistry

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The morphology and curvature of biological bilayers are determined by the packing shapes and interactions of their participant molecules. Bacteria, except photosynthetic groups, usually lack intracellular membrane organelles. Strong overexpression in Escherichia coli of a foreign monotopic glycosyltransferase (named monoglycosyldiacylglycerol synthase), synthesizing a nonbilayer-prone glucolipid, induced massive formation of membrane vesicles in the cytoplasm. Vesicle assemblies were visualized in cytoplasmic zones by fluorescence microscopy. These have a very low buoyant density, substantially different from inner membranes, with a lipid content of >60% (w/w). Cryo-transmission electron microscopy revealed cells to be filled with membrane vesicles of various sizes and shapes, which when released were mostly spherical (diameter ≈100 nm). The protein repertoire was similar in vesicle and inner membranes and dominated by the glycosyltransferase. Membrane polar lipid composition was similar too, including the foreign glucolipid. A related glycosyltransferase and an inactive monoglycosyldiacylglycerol synthase mutant also yielded membrane vesicles, but without glucolipid synthesis, strongly indicating that vesiculation is induced by the protein itself. The high capacity for membrane vesicle formation seems inherent in the glycosyltransferase structure, and it depends on the following: (i) lateral expansion of the inner monolayer by interface binding of many molecules; (ii) membrane expansion through stimulation of phospholipid synthesis, by electrostatic binding and sequestration of anionic lipids; (iii) bilayer bending by the packing shape of excess nonbilayer-prone phospholipid or glucolipid; and (iv) potentially also the shape or penetration profile of the glycosyltransferase binding surface. These features seem to apply to several other proteins able to achieve an analogous membrane expansion.

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

Cited by 73 publications

References 33 publications

Phospholipid Dependence and Liposome Reconstitution of Purified Hyaluronan Synthase

Phospholipid Dependence and Liposome Reconstitution of Purified Hyaluronan Synthase

Evolutionary genomics of ecological specialization

Massive Formation of Intracellular Membrane Vesicles in Escherichia coli by a Monotopic Membrane-bound Lipid Glycosyltransferase

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