Detergents modulate dimerization, but not helicity, of the glycophorin A transmembrane domain

Fisher, Lillian Estelle; Engelman, Donald M.; Sturgis, James N.

doi:10.1006/jmbi.1999.3126

Cited by 171 publications

(217 citation statements)

References 49 publications

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“…Mutagenesis studies have defined the amino acid motif involved in the interactions (Lemmon et al, 1992), and a reporter gene dimerization assay (TOXCAT) has confirmed this motif (Russ and Engelman, 1999). The interaction has also been quantified by FRET (Fisher et al, 1999). Final confirmation of these data are provided by the elucidation of the NMR structure of the GPA TM domain dimer (MacKenzie et al, 1997).…”

Section: Introductionmentioning

confidence: 91%

“…For the FRET experiments, protected and resin-bound peptides were chemically labeled with 1-pyrene butanoic acid (Molecular Probes, Eugene, OR) or dimethyl-amino-coumarin-acetic acid succinimidyl ester (DAMCA-SE; Fluoprobes, Interchim, Montluçon, France) in dimethylsulfoxide (Fisher et al, 1999). After de-protection, peptides were purified by RP-HPLC (Beckman Gold apparatus, Nucleosil C4 or Phenyl Macherey-Nagel (Hoerdt, France) semi-preparative columns.…”

Section: Labeling and Hplc Purification Of The Tm Peptidesmentioning

confidence: 99%

“…Reconstitution in detergent micelles and fluorescence resonance energy transfer (FRET) measurements were performed as previously described by Fisher et al (1999Fisher et al ( , 2003. Briefly, an equimolar mixture of pyrene-and coumarinlabeled peptides was adjusted to 5-20 M total peptides in trifluoroethanol (TFE) and was mixed with a small volume of 1 M lauryldimethylamine-oxide (LDAO; Fluka, L'Isle d'Abeau, France).…”

Section: Fluorescence Resonance Energy Transfer Measurementsmentioning

confidence: 99%

“…Fluorescence excitation spectra were recorded, and the relative contribution of pyrene (sensitized emission, with its characteristic peak at 345 nm) and coumarin (direct emission, with its characteristic peak at 370 nm) to the fluorescence emission at 500 nm was calculated. This FRET ratio provides a measure of the degree of dimerization of the two peptides (Fisher et al, 1999). An apparent dissociation constant was calculated from the dependence of the FRET signal on the peptide concentration, whereas the detergent concentration remained constant.…”

Section: Fluorescence Resonance Energy Transfer Measurementsmentioning

confidence: 99%

“…Furthermore, the Toxluc system revealed no binding capacity of the TM-NRP1 mut in which the three glycines of the GxxxG motif are replaced by valines. We also performed a FRET analysis of the TM NRP1 dimerization using the fluorescence assay developed by Fisher et al (1999), which permits the study of dimeric interactions. In this assay, fluorescently labeled synthetic peptides interact in detergent solution and the interaction is measured by the level of FRET between a donor and an acceptor fluorophore.…”

Section: Tm Nrp1 Has a Strong Dimerization Capacitymentioning

confidence: 99%

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Transmembrane Domain Interactions Control Biological Functions of Neuropilin-1

Roth

Nasarre

Dirrig‐Grosch

et al. 2008

MBoC

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Neuropilin-1 (NRP1) is a transmembrane receptor playing a pivotal role in the control of semaphorins and VEGF signaling pathways. The exact mechanism controlling semaphorin receptor complex formation is unknown. A structural analysis and modeling of NRP1 revealed a putative dimerization GxxxG motif potentially important for NRP1 dimerization and oligomerization. Our data show that this motif mediates the dimerization of the transmembrane domain of NRP1 as demonstrated by a dimerization assay (ToxLuc assay) performed in natural membrane and FRET analysis. A synthetic peptide derived from the transmembrane segment of NRP1 abolished the inhibitory effect of Sema3A. This effect depends on the capacity of the peptide to interfere with NRP1 dimerization and the formation of oligomeric complexes. Mutation of the GxxxG dimerization motif in the transmembrane domain of NRP1 confirmed its biological importance for Sema3A signaling. Overall, our results shed light on an essential step required for semaphorin signaling and provide novel evidence for the crucial role of transmembrane domain of bitopic protein containing GxxxG motif in the formation of receptor complexes that are a prerequisite for cell signaling.

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Section: Introductionmentioning

confidence: 91%

Section: Labeling and Hplc Purification Of The Tm Peptidesmentioning

confidence: 99%

Section: Fluorescence Resonance Energy Transfer Measurementsmentioning

confidence: 99%

Section: Fluorescence Resonance Energy Transfer Measurementsmentioning

confidence: 99%

Section: Tm Nrp1 Has a Strong Dimerization Capacitymentioning

confidence: 99%

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Transmembrane Domain Interactions Control Biological Functions of Neuropilin-1

Roth

Nasarre

Dirrig‐Grosch

et al. 2008

MBoC

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Glycophorin

Epand

2002

Wiley Encyclopedia of Molecular Medicine

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Transmemebrane Domains in Proteins

Ridder

Langosch

2008

Protein Science Encyclopedia

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Originally published in: Protein Folding Handbook. Part II. Edited by Johannes Buchner and Thomas Kiefhaber. Copyright © 2005 Wiley‐VCH Verlag GmbH & Co. KGaA Weinheim. Print ISBN: 3‐527‐30784‐2 The sections in this article are Introduction Structure of Transmembrane Domains The Biosynthetic Route towards Folded and Oligomeric Integral Membrane Proteins Structure and Stability of TMS s Amino Acid Composition of TMS s and Flanking Regions Stability of Transmembrane Helices The Nature of Transmembrane Helix‐Helix Interactions General Considerations Attractive Forces within Lipid Bilayers Forces between Transmembrane Helices Entropic Factors Influencing Transmembrane Helix‐Helix Interactions Lessons from Sequence Analyses and High‐resolution Structures Lessons from Bitopic Membrane Proteins Transmembrane Segments Forming Right‐handed Pairs Transmembrane Segments Forming Left‐handed Assemblies Selection of Self‐interacting TMS s from Combinatorial Libraries Role of Lipids in Packing/Assembly of Membrane Proteins Conformational Flexibility of Transmembrane Segments Experimental Techniques Biochemical and Biophysical Techniques Visualization of Oligomeric States by Electrophoretic Techniques Hydrodynamic Methods Fluorescence Resonance Transfer Genetic Assays The Tox R System Other Genetic Assays Identification of TMS ‐ TMS Interfaces by Mutational Analysis

show abstract

Detergents modulate dimerization, but not helicity, of the glycophorin A transmembrane domain

Cited by 171 publications

References 49 publications

Transmembrane Domain Interactions Control Biological Functions of Neuropilin-1

Transmembrane Domain Interactions Control Biological Functions of Neuropilin-1

Glycophorin

Transmemebrane Domains in Proteins

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