title: Effect of juxtamembrane tryptophans on the immersion depth of Synaptobrevin, an integral vesicle membrane protein.
abstract: Proper positioning of membrane proteins in the host membrane is often critical to successful protein function. While hydrophobic considerations play a dominant role in determining the topology of a protein in the membrane, amphiphilic residues, such as tryptophan, may 'anchor' the protein near the water-membrane interface. The SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) family of membrane proteins mediates intracellular membrane fusion. Correct positioning of the SNAREs is necessary if fusion is to occur. Synaptobrevins are integral vesicle membrane proteins that are well conserved across species. Interestingly, mammalian Synaptobrevins typically contain two adjacent tryptophans near the water-membrane interface whereas the Drosophila, neuronal-Synaptobrevin (n-Syb), contains a single tryptophan in this same region. To explore the role of these tryptophan residues in membrane positioning, we prepared a peptide containing residues 75-121 of D. melanogaster n-Syb in DPC micelles, biosynthetically labeled with 4-fluorophenylalanine and 5-fluorotryptophan for the examination by ¹⁹F NMR spectroscopy. Mutations of this construct containing zero and two tryptophan residues near the water-membrane interface resulted in changes in the positioning of n-Syb in the micelle. Moreover, the addition of a second tryptophan appears to slow dynamic motions of n-Syb near the micelle-water interface. These data therefore indicate that juxtamembrane tryptophan residues are important determinants of the position of Synaptobrevin in the membrane.
authors: Department of Chemistry, University of Toronto Mississauga, Ontario, Canada.
authors: M Sameer
authors: MS
authors: Al-Abdul-Wahid
authors: Colin M
authors: CM
authors: Demill
authors: Marzena B
authors: MB
authors: Serwin
authors: R Scott
authors: RS
authors: Prosser
authors: Bryan A
authors: BA
authors: Stewart
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D021961:Cell Membrane Structures; D004331:Drosophila melanogaster / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D008561:Membrane Fusion; D050683:R-SNARE Proteins / Q000737:chemistry* / Q000378:metabolism*; D014364:Tryptophan / Q000737:chemistry*
chemical_list: D050683:R-SNARE Proteins; D014364:Tryptophan