Logo of biochemjBJ Latest papers and much more!
Biochem J. 2004 Mar 15; 378(Pt 3): 1067–1072.
PMCID: PMC1224031

Phosphoinositides differentially regulate alpha-actinin flexibility and function.

Abstract

Alpha-actinin is a cell-adhesion and cytoskeletal protein that bundles actin microfilaments and links these filaments directly to integrin-adhesion receptors. Phosphoinositides bind to and regulate the interaction of a-actinin with actin filaments and integrin receptors. In the present study, we demonstrate that PtdIns(3,4,5)P3 inhibits and disrupts a-actinin-bundling activity, whereas PtdIns(4,5)P2 can only inhibit activity. In addition, a protease-sensitivity assay was developed to examine the flexibility of the linker region between the actin-binding domain and the spectrin repeats of a-actinin. Both phosphoinositides influenced the extent of proteolysis and the cleavage sites. PtdIns(4,5)P2 binding decreased the proteolysis of a-actinin, suggesting a role in stabilizing the structure of the protein. In contrast, PtdIns(3,4,5)P3 binding enhanced a-actinin proteolysis, indicating an increase in the flexibility of the protein. Furthermore, phosphoinositide binding influenced the proteolysis of the N- and C-terminal domains of a-actinin, indicating regulation of structure within both domains. These results support the hypothesis that PtdIns(4,5)P2 and PtdIns(3,4,5)P3 differentially regulate a-actinin function by modulating the structure and flexibility of the protein.

Full Text

The Full Text of this article is available as a PDF (225K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Otey CA, Pavalko FM, Burridge K. An interaction between alpha-actinin and the beta 1 integrin subunit in vitro. J Cell Biol. 1990 Aug;111(2):721–729. [PMC free article] [PubMed]
  • Blanchard A, Ohanian V, Critchley D. The structure and function of alpha-actinin. J Muscle Res Cell Motil. 1989 Aug;10(4):280–289. [PubMed]
  • Pavalko FM, Otey CA, Simon KO, Burridge K. Alpha-actinin: a direct link between actin and integrins. Biochem Soc Trans. 1991 Nov;19(4):1065–1069. [PubMed]
  • Tang J, Taylor DW, Taylor KA. The three-dimensional structure of alpha-actinin obtained by cryoelectron microscopy suggests a model for Ca(2+)-dependent actin binding. J Mol Biol. 2001 Jul 20;310(4):845–858. [PubMed]
  • Keep NH, Winder SJ, Moores CA, Walke S, Norwood FL, Kendrick-Jones J. Crystal structure of the actin-binding region of utrophin reveals a head-to-tail dimer. Structure. 1999 Dec 15;7(12):1539–1546. [PubMed]
  • Norwood FL, Sutherland-Smith AJ, Keep NH, Kendrick-Jones J. The structure of the N-terminal actin-binding domain of human dystrophin and how mutations in this domain may cause Duchenne or Becker muscular dystrophy. Structure. 2000 May 15;8(5):481–491. [PubMed]
  • Winkler J, Lünsdorf H, Jockusch BM. Flexibility and fine structure of smooth-muscle alpha-actinin. Eur J Biochem. 1997 Aug 15;248(1):193–199. [PubMed]
  • Young P, Gautel M. The interaction of titin and alpha-actinin is controlled by a phospholipid-regulated intramolecular pseudoligand mechanism. EMBO J. 2000 Dec 1;19(23):6331–6340. [PMC free article] [PubMed]
  • Fukami K, Sawada N, Endo T, Takenawa T. Identification of a phosphatidylinositol 4,5-bisphosphate-binding site in chicken skeletal muscle alpha-actinin. J Biol Chem. 1996 Feb 2;271(5):2646–2650. [PubMed]
  • Fraley Tamara S, Tran Thuan C, Corgan Anne Marie, Nash Coral A, Hao Jie, Critchley David R, Greenwood Jeffrey A. Phosphoinositide binding inhibits alpha-actinin bundling activity. J Biol Chem. 2003 Jun 27;278(26):24039–24045. [PubMed]
  • Greenwood JA, Theibert AB, Prestwich GD, Murphy-Ullrich JE. Restructuring of focal adhesion plaques by PI 3-kinase. Regulation by PtdIns (3,4,5)-p(3) binding to alpha-actinin. J Cell Biol. 2000 Aug 7;150(3):627–642. [PMC free article] [PubMed]
  • Pace CN, Vajdos F, Fee L, Grimsley G, Gray T. How to measure and predict the molar absorption coefficient of a protein. Protein Sci. 1995 Nov;4(11):2411–2423. [PMC free article] [PubMed]
  • Toumadje A, Johnson WC., Jr Effects of relative band intensity on prediction of protein secondary structure from CD. Anal Biochem. 1993 Jun;211(2):258–260. [PubMed]
  • Lu PJ, Chen CS. Selective recognition of phosphatidylinositol 3,4,5-trisphosphate by a synthetic peptide. J Biol Chem. 1997 Jan 3;272(1):466–472. [PubMed]
  • Lu PJ, Shieh WR, Rhee SG, Yin HL, Chen CS. Lipid products of phosphoinositide 3-kinase bind human profilin with high affinity. Biochemistry. 1996 Nov 5;35(44):14027–14034. [PubMed]
  • Imamura M, Endo T, Kuroda M, Tanaka T, Masaki T. Substructure and higher structure of chicken smooth muscle alpha-actinin molecule. J Biol Chem. 1988 Jun 5;263(16):7800–7805. [PubMed]
  • Manavalan P, Johnson WC., Jr Variable selection method improves the prediction of protein secondary structure from circular dichroism spectra. Anal Biochem. 1987 Nov 15;167(1):76–85. [PubMed]
  • Ching TT, Lin HP, Yang CC, Oliveira M, Lu PJ, Chen CS. Specific binding of the C-terminal Src homology 2 domain of the p85alpha subunit of phosphoinositide 3-kinase to phosphatidylinositol 3,4,5-trisphosphate. Localization and engineering of the phosphoinositide-binding motif. J Biol Chem. 2001 Nov 23;276(47):43932–43938. [PubMed]
  • Matsubara H, Singer A, Sasaki R, Jukes TH. Observations on the specificity of a thermostable bacterial protease "thermolysin". Biochem Biophys Res Commun. 1965 Nov 8;21(3):242–247. [PubMed]
  • Matsubara H, Sasaki R, Singer A, Jukes TH. Specific nature of hydrolysis of insulin and tobacco mosaic virus protein by thermolysin. Arch Biochem Biophys. 1966 Aug;115(2):324–331. [PubMed]
  • Bañuelos S, Saraste M, Djinović Carugo K. Structural comparisons of calponin homology domains: implications for actin binding. Structure. 1998 Nov 15;6(11):1419–1431. [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

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...