Monitoring pyrene excimers in lactose permease liposomes: revealing the presence of phosphatidylglycerol in proximity to an integral membrane protein

In this study, we examined the annular lipid composition of the transmembrane protein lactose permease (LacY) from Escherichia coli. LacY was reconstituted into 1-Palmitoyl-2-Oleoyl-sn-Glycero-3-Phosphoethanolamine (POPE) and 1-Palmitoyl-2-Oleoyl-sn-Glycero-3-3-[Phospho-rac-(1-glycerol)] (POPG) and...

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Bibliographic Details
Published in:Journal of fluorescence 2007-11, Vol.17 (6), p.649-654
Main Authors: Picas, Laura, Merino-Montero, Sandra, Morros, Antoni, Hernández-Borrell, Jordi, Montero, M Teresa
Format: Article
Language:English
Subjects:
Amino Acid Substitution
Biochemistry, Molecular Biology
Biophysics
Escherichia coli
Escherichia coli Proteins - chemistry
Escherichia coli Proteins - genetics
Hydrogen Bonding
Life Sciences
Liposomes
Microscopy, Atomic Force
Models, Molecular
Monosaccharide Transport Proteins - chemistry
Monosaccharide Transport Proteins - genetics
Phosphatidylglycerols - chemistry
Pyrenes - chemistry
Spectrophotometry
Symporters - chemistry
Symporters - genetics
Thermodynamics
Tryptophan - chemistry
Tryptophan - genetics
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Summary:In this study, we examined the annular lipid composition of the transmembrane protein lactose permease (LacY) from Escherichia coli. LacY was reconstituted into 1-Palmitoyl-2-Oleoyl-sn-Glycero-3-Phosphoethanolamine (POPE) and 1-Palmitoyl-2-Oleoyl-sn-Glycero-3-3-[Phospho-rac-(1-glycerol)] (POPG) and labeled with 1-hexadecanoyl-2-(1-pyrenedecanoyl)-sn-Glycero-3-phosphoglycerol (PPDPG) at a 3:0.99:0.01 molar ratio. Pyrene excimer formation was monitored by exciting a single tryptophan mutant of the protein (T320W). The results suggest that POPG remains segregated in the vicinity of the protein, most likely forming part of the annular composition. The possible involvement of POPG in hydrogen binding with the protein, as well as the molecular mechanism of LacY, is also discussed in the context of the proteomic network theory.
ISSN:1053-0509
1573-4994
DOI:10.1007/s10895-006-0073-0