PMID:11732909

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Citation

Pilot, JD, East, JM and Lee, AG (2001) Effects of phospholipid headgroup and phase on the activity of diacylglycerol kinase of Escherichia coli. Biochemistry 40:14891-7

Abstract

Diacylglycerol kinase (DGK) of Escherichia coli has been reconstituted into a variety of phospholipid bilayers and its activity determined as a function of lipid headgroup structure and phase preference. The anionic phospholipids dioleoylphosphatidic acid, dioleoylphosphatidylserine, and cardiolipin were all found to support activities lower than that supported by dioleoylphosphatidylcholine. In mixtures of dioleoylphosphatidylcholine and 20 mol % anionic phospholipids, the presence of anionic phospholipids all resulted in lower activities than in dioleoylphosphatidylcholine, except for dioleoylphosphatidylglycerol whose presence had little effect on activity. In some cases, the low activity in the presence of anionic phospholipid followed from a decrease in v(max); in some cases, it followed from an increase in the K(m) for diacylglycerol, and in the case of dioleoylphosphatidic acid, it followed from both. Activities in mixtures containing 80 mol % dioleoylphosphatidylethanolamine were lower than in dioleoylphosphatidylcholine at temperatures where both lipids adopted a bilayer phase; at higher temperatures where dioleoylphosphatidylethanolamine preferred a hexagonal H(II) phase, the differences in activity were greater. These experiments suggest that the presence of lipids preferring a hexagonal H(II) phase leads to low activities. Activities of DGK are low in a gel phase lipid.

Links

PubMed

Keywords

Cell Membrane/chemistry; Cell Membrane/metabolism; Diacylglycerol Kinase/metabolism; Escherichia coli/chemistry; Escherichia coli/enzymology; Escherichia coli/metabolism; Lipid Bilayers/chemistry; Lipid Bilayers/metabolism; Magnesium/metabolism; Phospholipids/chemistry; Phospholipids/metabolism; Temperature

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