• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of embojLink to Publisher's site
EMBO J. Feb 3, 1997; 16(3): 484–494.
PMCID: PMC1169652

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.

Full Text

The Full Text of this article is available as a PDF (559K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ampe C, Markey F, Lindberg U, Vandekerckhove J. The primary structure of human platelet profilin: reinvestigation of the calf spleen profilin sequence. FEBS Lett. 1988 Feb 8;228(1):17–21. [PubMed]
  • Archer SJ, Vinson VK, Pollard TD, Torchia DA. Elucidation of the poly-L-proline binding site in Acanthamoeba profilin I by NMR spectroscopy. FEBS Lett. 1994 Jan 10;337(2):145–151. [PubMed]
  • Björkegren C, Rozycki M, Schutt CE, Lindberg U, Karlsson R. Mutagenesis of human profilin locates its poly(L-proline)-binding site to a hydrophobic patch of aromatic amino acids. FEBS Lett. 1993 Oct 25;333(1-2):123–126. [PubMed]
  • Buss F, Jockusch BM. Tissue-specific expression of profilin. FEBS Lett. 1989 May 22;249(1):31–34. [PubMed]
  • Buss F, Temm-Grove C, Henning S, Jockusch BM. Distribution of profilin in fibroblasts correlates with the presence of highly dynamic actin filaments. Cell Motil Cytoskeleton. 1992;22(1):51–61. [PubMed]
  • Lassing I, Lindberg U. Specificity of the interaction between phosphatidylinositol 4,5-bisphosphate and the profilin:actin complex. J Cell Biochem. 1988 Jul;37(3):255–267. [PubMed]
  • Carlsson L, Nyström LE, Sundkvist I, Markey F, Lindberg U. Actin polymerizability is influenced by profilin, a low molecular weight protein in non-muscle cells. J Mol Biol. 1977 Sep 25;115(3):465–483. [PubMed]
  • Machesky LM, Cole NB, Moss B, Pollard TD. Vaccinia virus expresses a novel profilin with a higher affinity for polyphosphoinositides than actin. Biochemistry. 1994 Sep 6;33(35):10815–10824. [PubMed]
  • Domann E, Wehland J, Rohde M, Pistor S, Hartl M, Goebel W, Leimeister-Wächter M, Wuenscher M, Chakraborty T. A novel bacterial virulence gene in Listeria monocytogenes required for host cell microfilament interaction with homology to the proline-rich region of vinculin. EMBO J. 1992 May;11(5):1981–1990. [PMC free article] [PubMed]
  • Metzler WJ, Bell AJ, Ernst E, Lavoie TB, Mueller L. Identification of the poly-L-proline-binding site on human profilin. J Biol Chem. 1994 Feb 11;269(6):4620–4625. [PubMed]
  • Metzler WJ, Farmer BT, 2nd, Constantine KL, Friedrichs MS, Lavoie T, Mueller L. Refined solution structure of human profilin I. Protein Sci. 1995 Mar;4(3):450–459. [PMC free article] [PubMed]
  • Niebuhr K, Chakraborty T, Rohde M, Gazlig T, Jansen B, Köllner P, Wehland J. Localization of the ActA polypeptide of Listeria monocytogenes in infected tissue culture cell lines: ActA is not associated with actin "comets". Infect Immun. 1993 Jul;61(7):2793–2802. [PMC free article] [PubMed]
  • Gerst JE, Ferguson K, Vojtek A, Wigler M, Field J. CAP is a bifunctional component of the Saccharomyces cerevisiae adenylyl cyclase complex. Mol Cell Biol. 1991 Mar;11(3):1248–1257. [PMC free article] [PubMed]
  • Pantaloni D, Carlier MF. How profilin promotes actin filament assembly in the presence of thymosin beta 4. Cell. 1993 Dec 3;75(5):1007–1014. [PubMed]
  • Goldschmidt-Clermont PJ, Janmey PA. Profilin, a weak CAP for actin and RAS. Cell. 1991 Aug 9;66(3):419–421. [PubMed]
  • Perelroizen I, Marchand JB, Blanchoin L, Didry D, Carlier MF. Interaction of profilin with G-actin and poly(L-proline). Biochemistry. 1994 Jul 19;33(28):8472–8478. [PubMed]
  • Ponder JW, Richards FM. Tertiary templates for proteins. Use of packing criteria in the enumeration of allowed sequences for different structural classes. J Mol Biol. 1987 Feb 20;193(4):775–791. [PubMed]
  • Goldschmidt-Clermont PJ, Kim JW, Machesky LM, Rhee SG, Pollard TD. Regulation of phospholipase C-gamma 1 by profilin and tyrosine phosphorylation. Science. 1991 Mar 8;251(4998):1231–1233. [PubMed]
  • Haffner C, Jarchau T, Reinhard M, Hoppe J, Lohmann SM, Walter U. Molecular cloning, structural analysis and functional expression of the proline-rich focal adhesion and microfilament-associated protein VASP. EMBO J. 1995 Jan 3;14(1):19–27. [PMC free article] [PubMed]
  • Schutt CE, Myslik JC, Rozycki MD, Goonesekere NC, Lindberg U. The structure of crystalline profilin-beta-actin. Nature. 1993 Oct 28;365(6449):810–816. [PubMed]
  • Halbrügge M, Friedrich C, Eigenthaler M, Schanzenbächer P, Walter U. Stoichiometric and reversible phosphorylation of a 46-kDa protein in human platelets in response to cGMP- and cAMP-elevating vasodilators. J Biol Chem. 1990 Feb 25;265(6):3088–3093. [PubMed]
  • Sohn RH, Chen J, Koblan KS, Bray PF, Goldschmidt-Clermont PJ. Localization of a binding site for phosphatidylinositol 4,5-bisphosphate on human profilin. J Biol Chem. 1995 Sep 8;270(36):21114–21120. [PubMed]
  • Honoré B, Madsen P, Andersen AH, Leffers H. Cloning and expression of a novel human profilin variant, profilin II. FEBS Lett. 1993 Sep 13;330(2):151–155. [PubMed]
  • Tanaka M, Shibata H. Poly(L-proline)-binding proteins from chick embryos are a profilin and a profilactin. Eur J Biochem. 1985 Sep 2;151(2):291–297. [PubMed]
  • Kaiser DA, Goldschmidt-Clermont PJ, Levine BA, Pollard TD. Characterization of renatured profilin purified by urea elution from poly-L-proline agarose columns. Cell Motil Cytoskeleton. 1989;14(2):251–262. [PubMed]
  • Theriot JA, Rosenblatt J, Portnoy DA, Goldschmidt-Clermont PJ, Mitchison TJ. Involvement of profilin in the actin-based motility of L. monocytogenes in cells and in cell-free extracts. Cell. 1994 Feb 11;76(3):505–517. [PubMed]
  • Turunen O, Wahlström T, Vaheri A. Ezrin has a COOH-terminal actin-binding site that is conserved in the ezrin protein family. J Cell Biol. 1994 Sep;126(6):1445–1453. [PMC free article] [PubMed]
  • Vojtek A, Haarer B, Field J, Gerst J, Pollard TD, Brown S, Wigler M. Evidence for a functional link between profilin and CAP in the yeast S. cerevisiae. Cell. 1991 Aug 9;66(3):497–505. [PubMed]
  • Waldmann R, Bauer S, Göbel C, Hofmann F, Jakobs KH, Walter U. Demonstration of cGMP-dependent protein kinase and cGMP-dependent phosphorylation in cell-free extracts of platelets. Eur J Biochem. 1986 Jul 1;158(1):203–210. [PubMed]
  • Waldmann R, Nieberding M, Walter U. Vasodilator-stimulated protein phosphorylation in platelets is mediated by cAMP- and cGMP-dependent protein kinases. Eur J Biochem. 1987 Sep 15;167(3):441–448. [PubMed]
  • Zeile WL, Purich DL, Southwick FS. Recognition of two classes of oligoproline sequences in profilin-mediated acceleration of actin-based Shigella motility. J Cell Biol. 1996 Apr;133(1):49–59. [PMC free article] [PubMed]

Articles from The EMBO Journal are provided here courtesy of The European Molecular Biology Organization

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...