Characterization of Two Classes of Small Molecule Inhibitors of Arp2/3 Complex

Overview

Journal Nature

Specialty Science

Date 2009 Aug 4

PMID 19648907

Citations 309

Authors

B J Nolen

N Tomasevic

A Russell

D W Pierce

Z Jia

C D McCormick

J Hartman

R Sakowicz

T D Pollard

Affiliations

Soon will be listed here.

Abstract

Polymerization of actin filaments directed by the actin-related protein (Arp)2/3 complex supports many types of cellular movements. However, questions remain regarding the relative contributions of Arp2/3 complex versus other mechanisms of actin filament nucleation to processes such as path finding by neuronal growth cones; this is because of the lack of simple methods to inhibit Arp2/3 complex reversibly in living cells. Here we describe two classes of small molecules that bind to different sites on the Arp2/3 complex and inhibit its ability to nucleate actin filaments. CK-0944636 binds between Arp2 and Arp3, where it appears to block movement of Arp2 and Arp3 into their active conformation. CK-0993548 inserts into the hydrophobic core of Arp3 and alters its conformation. Both classes of compounds inhibit formation of actin filament comet tails by Listeria and podosomes by monocytes. Two inhibitors with different mechanisms of action provide a powerful approach for studying the Arp2/3 complex in living cells.

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References

Blanchoin L, Amann K, Higgs H, Marchand J, Kaiser D, Pollard T . Direct observation of dendritic actin filament networks nucleated by Arp2/3 complex and WASP/Scar proteins. Nature. 2000; 404(6781):1007-11. DOI: 10.1038/35010008. View

Jabs A, Weiss M, Hilgenfeld R . Non-proline cis peptide bonds in proteins. J Mol Biol. 1999; 286(1):291-304. DOI: 10.1006/jmbi.1998.2459. View

Goley E, Welch M . The ARP2/3 complex: an actin nucleator comes of age. Nat Rev Mol Cell Biol. 2006; 7(10):713-26. DOI: 10.1038/nrm2026. View

Lee J, Ishihara A, Theriot J, Jacobson K . Principles of locomotion for simple-shaped cells. Nature. 1993; 362(6416):167-71. DOI: 10.1038/362167a0. View

Holmes K, Popp D, GEBHARD W, Kabsch W . Atomic model of the actin filament. Nature. 1990; 347(6288):44-9. DOI: 10.1038/347044a0. View

Gimona M, Buccione R, Courtneidge S, Linder S . Assembly and biological role of podosomes and invadopodia. Curr Opin Cell Biol. 2008; 20(2):235-41. DOI: 10.1016/j.ceb.2008.01.005. View

Nolen B, Pollard T . Structure and biochemical properties of fission yeast Arp2/3 complex lacking the Arp2 subunit. J Biol Chem. 2008; 283(39):26490-8. PMC: 2546537. DOI: 10.1074/jbc.M802607200. View

Linder S, Higgs H, Hufner K, Schwarz K, Pannicke U, Aepfelbacher M . The polarization defect of Wiskott-Aldrich syndrome macrophages is linked to dislocalization of the Arp2/3 complex. J Immunol. 2000; 165(1):221-5. DOI: 10.4049/jimmunol.165.1.221. View

Cooper J, WALKER S, Pollard T . Pyrene actin: documentation of the validity of a sensitive assay for actin polymerization. J Muscle Res Cell Motil. 1983; 4(2):253-62. DOI: 10.1007/BF00712034. View

10.

Nolen B, Littlefield R, Pollard T . Crystal structures of actin-related protein 2/3 complex with bound ATP or ADP. Proc Natl Acad Sci U S A. 2004; 101(44):15627-32. PMC: 524860. DOI: 10.1073/pnas.0407149101. View

11.

Pollard T . Identification of a factor in conventional muscle actin preparations which inhibits actin filament self-association. Biochem Biophys Res Commun. 1980; 96(1):18-27. DOI: 10.1016/0006-291x(80)91175-4. View

12.

Kovar D, Pollard T . Insertional assembly of actin filament barbed ends in association with formins produces piconewton forces. Proc Natl Acad Sci U S A. 2004; 101(41):14725-30. PMC: 522035. DOI: 10.1073/pnas.0405902101. View

13.

Dodson E, Winn M, Ralph A . Collaborative Computational Project, number 4: providing programs for protein crystallography. Methods Enzymol. 1997; 277:620-33. DOI: 10.1016/s0076-6879(97)77034-4. View

14.

Dayel M, Holleran E, Mullins R . Arp2/3 complex requires hydrolyzable ATP for nucleation of new actin filaments. Proc Natl Acad Sci U S A. 2001; 98(26):14871-6. PMC: 64951. DOI: 10.1073/pnas.261419298. View

15.

Higgs H, Blanchoin L, Pollard T . Influence of the C terminus of Wiskott-Aldrich syndrome protein (WASp) and the Arp2/3 complex on actin polymerization. Biochemistry. 1999; 38(46):15212-22. DOI: 10.1021/bi991843+. View

16.

Brunger A, Adams P, Clore G, DeLano W, Gros P, Grosse-Kunstleve R . Crystallography & NMR system: A new software suite for macromolecular structure determination. Acta Crystallogr D Biol Crystallogr. 1998; 54(Pt 5):905-21. DOI: 10.1107/s0907444998003254. View

17.

Schuttelkopf A, van Aalten D . PRODRG: a tool for high-throughput crystallography of protein-ligand complexes. Acta Crystallogr D Biol Crystallogr. 2004; 60(Pt 8):1355-63. DOI: 10.1107/S0907444904011679. View

18.

Robinson R, Turbedsky K, Kaiser D, Marchand J, Higgs H, Choe S . Crystal structure of Arp2/3 complex. Science. 2001; 294(5547):1679-84. DOI: 10.1126/science.1066333. View

19.

Otwinowski Z, Minor W . Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 1997; 276:307-26. DOI: 10.1016/S0076-6879(97)76066-X. View

20.

Pollard T . Polymerization of ADP-actin. J Cell Biol. 1984; 99(3):769-77. PMC: 2113399. DOI: 10.1083/jcb.99.3.769. View