title: Co-operation of domain-binding and calcium-binding sites in the activation of gelsolin.
abstract: Gelsolin is an abundant calcium dependent actin filament severing and capping protein. In the absence of calcium the molecule is compact but in the presence of calcium, as its six similar domains alter their relative position, a generally more open configuration is adopted to reveal the three actin binding sites. It is generally held that a 'helical-latch' at the C-terminus of gelsolin's domain 6 (G6), binds domain 2 (G2) to keep gelsolin in the calcium-free compact state, and that the crutial calcium binding site(s) reside in the C-terminal half of gelsolin perhaps involving the C-terminal helix itself has to be bound to release this latch. Here we provide evidence for a calcium dependent conformational change within G2 (Kd = approximately 15 micro m). We also report a calcium dependent binding site for the C-terminus (G4-6) within G2 and delimit this further to a specific region formed by residues 203-225 and 159-193. It is known that the activation of gelsolin involves multiple calcium binding events (around 6) the first of which (in G6) may release the latch. We propose that the calcium-dependent conformational change in G2 may be a subsequent step that is necessary for the dissociation of G2 from G4-6, and that this movement occurs in sympathy with calcium induced conformational changes within G6 by the physical coupling of the two calcium binding sites within G2 and G6. Additional calcium binding in other domains then result in the complete opening and activation of the gelsolin molecule.
authors.affiliation: UMR 5539 (CNRS), Laboratoire de motilité cellulaire (Ecole Pratique des Hautes Etudes), Université de Montpellier 2, France.
authors.forename: Emeline
authors.initials: E
authors.lastname: Lagarrigue
authors.forename: Sutherland K
authors.initials: SK
authors.lastname: Maciver
authors.forename: Abdellatif
authors.initials: A
authors.lastname: Fattoum
authors.forename: Yves
authors.initials: Y
authors.lastname: Benyamin
authors.forename: Claude
authors.initials: C
authors.lastname: Roustan
mesh_terms: D000199:Actins / Q000737:chemistry; D001665:Binding Sites; D002118:Calcium / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D003599:Cytoskeleton / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D018260:Gelsolin / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence; D014357:Trypsin / Q000494:pharmacology
chemical_list: D000199:Actins; D018260:Gelsolin; D010455:Peptides; D014357:Trypsin; D002118:Calcium