title: Phospholipid binding of synthetic talin peptides provides evidence for an intrinsic membrane anchor of talin.
abstract: Talin, an actin-binding protein, is assumed to anchor at the membrane via an intrinsic amino acid sequence. Three N-terminal talin fragments, 21-39 (S19), 287-304 (H18), and 385-406 (H17) have been proposed as potential membrane anchors. The interaction of the corresponding synthetic peptides with lipid model systems was investigated with CD spectroscopy, isothermal titration calorimetry, and monolayer expansion measurements. The membrane model systems were neutral or negatively charged small unilamellar vesicles or monolayers with a lateral packing density of bilayers (32 mN/m). S19 partitions into charged monolayers/bilayers with a penetration area A(p) = 140 +/- 30 A(2) and a free energy of binding of DeltaG(0) = -5.7 kcal/mol, thereby forming a partially alpha-helical structure. H18 does not interact with lipid monolayers or bilayers. H17 penetrates into neutral and charged monolayers/bilayers with A(p) = 148 +/- 23 A(2) and A(p) = 160 +/- 15 A(2), respectively, forming an alpha-helix in the membrane-bound state. Membrane partitioning is mainly entropy-driven. Under physiological conditions the free energy of binding to negatively charged membranes is DeltaG(0) = -9. 4 kcal/mol with a hydrophobic contribution of DeltaG(h) = -7.8 kcal/mol, comparable to that of post-translationally attached membrane anchors, and an electrostatic contribution of DeltaG(h) = -1.6 kcal/mol. The latter becomes more negative with decreasing pH. We show that H17 provides the binding energy required for a membrane anchor.
authors.affiliation: Department of Biophysical Chemistry, Biocenter, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland. anna.seelig@unibas.ch
authors.forename: A
authors.initials: A
authors.lastname: Seelig
authors.forename: X L
authors.initials: XL
authors.lastname: Blatter
authors.forename: A
authors.initials: A
authors.lastname: Frentzel
authors.forename: G
authors.initials: G
authors.lastname: Isenberg
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002151:Calorimetry; D002462:Cell Membrane / Q000378:metabolism; D002942:Circular Dichroism; D007700:Kinetics; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000378:metabolism*; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D055672:Static Electricity; D013329:Structure-Activity Relationship; D016608:Talin / Q000378:metabolism*
chemical_list: D010743:Phospholipids; D016608:Talin