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Monitoring Pyrene Excimers in Lactose Permease Liposomes: Revealing the Presence of Phosphatidylglycerol in Proximity to an Integral Membrane Protein

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Abstract

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.

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Notes

  1. The one-letter amino acid code is used.

  2. The value for the POPE:POPG containing X PPDPG=0.01 was T m=18.19°C; the width at half-height of the thermogram 7.81°C; and the enthalpy change of the endotherm measured 13.90 kJ·mol−1.

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Acknowledgments

L. P. is the recipient of a collaborative fellowship from the Ministerio de Educación y Ciencia, and a S. M-M. is a fellow of 'Recerca i Docència' from the Universitat de Barcelona. We would like to thank Professor Ronald Kaback for the materials provided, and Dr. José Luis Vázquez-Ibar for engineering the single-Trp mutant, as well as for helpful comments on the original manuscript. This work was supported by grants CTQ 2005-07989 from the Ministerio de Educación y Ciencia, Spain.

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Authors and Affiliations

  1. Departament de Fisicoquímica, Facultat de Farmàcia, Universitat de Barcelona, Barcelona, 08028, Spain

    Laura Picas, Sandra Merino-Montero, Jordi Hernández-Borrell & M. Teresa Montero

  2. Unitat de Biofísica, Departament de Bioquímica i Biología Molecular, Facultat de Medicina, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain

    Antoni Morros

  3. Centre de Bioelectrònica i Nanobiociència (CBEN), Parc Científic de Barcelona, Barcelona, 08028, Spain

    Sandra Merino-Montero, Jordi Hernández-Borrell & M. Teresa Montero

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  1. Laura Picas
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  2. Sandra Merino-Montero
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  5. M. Teresa Montero
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Correspondence to M. Teresa Montero.

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Picas, L., Merino-Montero, S., Morros, A. et al. Monitoring Pyrene Excimers in Lactose Permease Liposomes: Revealing the Presence of Phosphatidylglycerol in Proximity to an Integral Membrane Protein. J Fluoresc 17, 649–654 (2007). https://doi.org/10.1007/s10895-006-0073-0

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  • DOI: https://doi.org/10.1007/s10895-006-0073-0

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