Dependence of M13 Major Coat Protein Oligomerization and Lateral Segregation of Bilayer Composition

F. Fernandes, L.M.S. Loura, M. Prieto, R.B.M. Koehorst, R.B. Spruijt, M.A. Hemminga

Research output: Contribution to journalArticleAcademicpeer-review

35 Citations (Scopus)

Abstract

M13 major coat protein was derivatized with BODIPY (n-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl iodoacetamide), and its aggregation was studied in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and DOPC/1,2-dioleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DOPG) or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)/DOPG (model systems of membranes with hydrophobic thickness matching that of the protein) using photophysical methodologies (time-resolved and steady-state self-quenching, absorption, and emission spectra). It was concluded that the protein is essentially monomeric, even in the absence of anionic phospholipids. The protein was also incorporated in pure bilayers of lipids with a strong mismatch with the protein transmembrane length, 1,2-dierucoyl-sn-glycero-3-phosphocholine (DEuPC, longer lipid) and 1,2-dimyristoleoyl-sn-glycero-3-phosphocholine (DMoPC, shorter lipid), and in lipidic mixtures containing DOPC and one of these lipids. The protein was aggregated in the pure vesicles of mismatching lipid but remained essentially monomeric in the mixtures as detected from BODIPY fluorescence emission self-quenching. From fluorescence resonance energy transfer (FRET) measurements (donor-n-(iodoacetyl)aminoethyl-1-sulfonaphthylamine (IAEDANS)-labeled protein; acceptor-BODIPY labeled protein), it was concluded that in the DEuPC/DOPC and DMoPC/DOPC lipid mixtures, domains enriched in the protein and the matching lipid (DOPC) are formed.
Original languageEnglish
Pages (from-to)2430-2441
Number of pages12
JournalBiophysical Journal
Volume85
Issue number4
DOIs
Publication statusPublished - 2003

Fingerprint

Capsid Proteins
Phosphorylcholine
Lipids
Proteins
Iodoacetamide
Fluorescence Resonance Energy Transfer
Lipid Bilayers
Phospholipids
Fluorescence
Membranes
4,4-difluoro-4-bora-3a,4a-diaza-s-indacene
1,2-dioleoyl-sn-glycero-3-phosphoglycerol

Keywords

  • dipyrrometheneboron difluoride bodipy
  • channel-associated peptide
  • resonance energy-transfer
  • lipid-bilayers
  • escherichia-coli
  • model membranes
  • hydrophobic mismatch
  • transmembrane domain
  • cytoplasmic membrane
  • diffusion

Cite this

Fernandes, F. ; Loura, L.M.S. ; Prieto, M. ; Koehorst, R.B.M. ; Spruijt, R.B. ; Hemminga, M.A. / Dependence of M13 Major Coat Protein Oligomerization and Lateral Segregation of Bilayer Composition. In: Biophysical Journal. 2003 ; Vol. 85, No. 4. pp. 2430-2441.
@article{aae18cac5f3041ffb687cef94a44aaa6,
title = "Dependence of M13 Major Coat Protein Oligomerization and Lateral Segregation of Bilayer Composition",
abstract = "M13 major coat protein was derivatized with BODIPY (n-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl iodoacetamide), and its aggregation was studied in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and DOPC/1,2-dioleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DOPG) or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)/DOPG (model systems of membranes with hydrophobic thickness matching that of the protein) using photophysical methodologies (time-resolved and steady-state self-quenching, absorption, and emission spectra). It was concluded that the protein is essentially monomeric, even in the absence of anionic phospholipids. The protein was also incorporated in pure bilayers of lipids with a strong mismatch with the protein transmembrane length, 1,2-dierucoyl-sn-glycero-3-phosphocholine (DEuPC, longer lipid) and 1,2-dimyristoleoyl-sn-glycero-3-phosphocholine (DMoPC, shorter lipid), and in lipidic mixtures containing DOPC and one of these lipids. The protein was aggregated in the pure vesicles of mismatching lipid but remained essentially monomeric in the mixtures as detected from BODIPY fluorescence emission self-quenching. From fluorescence resonance energy transfer (FRET) measurements (donor-n-(iodoacetyl)aminoethyl-1-sulfonaphthylamine (IAEDANS)-labeled protein; acceptor-BODIPY labeled protein), it was concluded that in the DEuPC/DOPC and DMoPC/DOPC lipid mixtures, domains enriched in the protein and the matching lipid (DOPC) are formed.",
keywords = "dipyrrometheneboron difluoride bodipy, channel-associated peptide, resonance energy-transfer, lipid-bilayers, escherichia-coli, model membranes, hydrophobic mismatch, transmembrane domain, cytoplasmic membrane, diffusion",
author = "F. Fernandes and L.M.S. Loura and M. Prieto and R.B.M. Koehorst and R.B. Spruijt and M.A. Hemminga",
year = "2003",
doi = "10.1016/S0006-3495(03)74666-9",
language = "English",
volume = "85",
pages = "2430--2441",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Biophysical Society",
number = "4",

}

Dependence of M13 Major Coat Protein Oligomerization and Lateral Segregation of Bilayer Composition. / Fernandes, F.; Loura, L.M.S.; Prieto, M.; Koehorst, R.B.M.; Spruijt, R.B.; Hemminga, M.A.

In: Biophysical Journal, Vol. 85, No. 4, 2003, p. 2430-2441.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Dependence of M13 Major Coat Protein Oligomerization and Lateral Segregation of Bilayer Composition

AU - Fernandes, F.

AU - Loura, L.M.S.

AU - Prieto, M.

AU - Koehorst, R.B.M.

AU - Spruijt, R.B.

AU - Hemminga, M.A.

PY - 2003

Y1 - 2003

N2 - M13 major coat protein was derivatized with BODIPY (n-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl iodoacetamide), and its aggregation was studied in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and DOPC/1,2-dioleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DOPG) or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)/DOPG (model systems of membranes with hydrophobic thickness matching that of the protein) using photophysical methodologies (time-resolved and steady-state self-quenching, absorption, and emission spectra). It was concluded that the protein is essentially monomeric, even in the absence of anionic phospholipids. The protein was also incorporated in pure bilayers of lipids with a strong mismatch with the protein transmembrane length, 1,2-dierucoyl-sn-glycero-3-phosphocholine (DEuPC, longer lipid) and 1,2-dimyristoleoyl-sn-glycero-3-phosphocholine (DMoPC, shorter lipid), and in lipidic mixtures containing DOPC and one of these lipids. The protein was aggregated in the pure vesicles of mismatching lipid but remained essentially monomeric in the mixtures as detected from BODIPY fluorescence emission self-quenching. From fluorescence resonance energy transfer (FRET) measurements (donor-n-(iodoacetyl)aminoethyl-1-sulfonaphthylamine (IAEDANS)-labeled protein; acceptor-BODIPY labeled protein), it was concluded that in the DEuPC/DOPC and DMoPC/DOPC lipid mixtures, domains enriched in the protein and the matching lipid (DOPC) are formed.

AB - M13 major coat protein was derivatized with BODIPY (n-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-yl)methyl iodoacetamide), and its aggregation was studied in 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and DOPC/1,2-dioleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DOPG) or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)/DOPG (model systems of membranes with hydrophobic thickness matching that of the protein) using photophysical methodologies (time-resolved and steady-state self-quenching, absorption, and emission spectra). It was concluded that the protein is essentially monomeric, even in the absence of anionic phospholipids. The protein was also incorporated in pure bilayers of lipids with a strong mismatch with the protein transmembrane length, 1,2-dierucoyl-sn-glycero-3-phosphocholine (DEuPC, longer lipid) and 1,2-dimyristoleoyl-sn-glycero-3-phosphocholine (DMoPC, shorter lipid), and in lipidic mixtures containing DOPC and one of these lipids. The protein was aggregated in the pure vesicles of mismatching lipid but remained essentially monomeric in the mixtures as detected from BODIPY fluorescence emission self-quenching. From fluorescence resonance energy transfer (FRET) measurements (donor-n-(iodoacetyl)aminoethyl-1-sulfonaphthylamine (IAEDANS)-labeled protein; acceptor-BODIPY labeled protein), it was concluded that in the DEuPC/DOPC and DMoPC/DOPC lipid mixtures, domains enriched in the protein and the matching lipid (DOPC) are formed.

KW - dipyrrometheneboron difluoride bodipy

KW - channel-associated peptide

KW - resonance energy-transfer

KW - lipid-bilayers

KW - escherichia-coli

KW - model membranes

KW - hydrophobic mismatch

KW - transmembrane domain

KW - cytoplasmic membrane

KW - diffusion

U2 - 10.1016/S0006-3495(03)74666-9

DO - 10.1016/S0006-3495(03)74666-9

M3 - Article

VL - 85

SP - 2430

EP - 2441

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 4

ER -