title: The mammalian profilin isoforms display complementary affinities for PIP2 and proline-rich sequences.
abstract: We present a study on the binding properties of the bovine profilin isoforms to both phosphatidylinositol 4,5-bisphosphate (PIP2) and proline-rich peptides derived from vasodilator-stimulated phosphoprotein (VASP) and cyclase-associated protein (CAP). Using microfiltration, we show that compared with profilin II, profilin I has a higher affinity for PIP2. On the other hand, fluorescence spectroscopy reveals that proline-rich peptides bind better to profilin II. At micromolar concentrations, profilin II dimerizes upon binding to proline-rich peptides. Circular dichroism measurements of profilin II reveal a significant conformational change in this protein upon binding of the peptide. We show further that PIP2 effectively competes for binding of profilin I to poly-L-proline, since this isoform, but not profilin II, can be eluted from a poly-L-proline column with PIP2. Using affinity chromatography on either profilin isoform, we identified profilin II as the preferred ligand for VASP in bovine brain extracts. The complementary affinities of the profilin isoforms for PIP2 and the proline-rich peptides offer the cell an opportunity to direct actin assembly at different subcellular localizations through the same or different signal transduction pathways.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Universiteit Gent, Belgium.
authors.forename: A
authors.initials: A
authors.lastname: Lambrechts
authors.forename: J L
authors.initials: JL
authors.lastname: Verschelde
authors.forename: V
authors.initials: V
authors.lastname: Jonckheere
authors.forename: M
authors.initials: M
authors.lastname: Goethals
authors.forename: J
authors.initials: J
authors.lastname: Vandekerckhove
authors.forename: C
authors.initials: C
authors.lastname: Ampe
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D002417:Cattle; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D018797:Cell Cycle Proteins; D002942:Circular Dichroism; D003285:Contractile Proteins; D003598:Cytoskeletal Proteins; D019281:Dimerization; D029721:Drosophila Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000378:metabolism*; D008840:Microfilament Proteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D051304:Profilins; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D013154:Spleen / Q000378:metabolism; D000098662:Vasodilator-Stimulated Phosphoprotein
chemical_list: C410502:Capt protein, Drosophila; D015815:Cell Adhesion Molecules; D018797:Cell Cycle Proteins; D003285:Contractile Proteins; D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins; D005656:Fungal Proteins; D008840:Microfilament Proteins; D010446:Peptide Fragments; D010455:Peptides; D019269:Phosphatidylinositol 4,5-Diphosphate; D010750:Phosphoproteins; D051304:Profilins; D000098662:Vasodilator-Stimulated Phosphoprotein; C011083:polyproline