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PDBsum entry 1t84

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Signaling protein PDB id
1t84
Jmol
Contents
Protein chain
107 a.a. *
Ligands
WSK
* Residue conservation analysis
PDB id:
1t84
Name: Signaling protein
Title: Solution structure of the wiskott-aldrich syndrome protein (wasp) autoinhibited core domain complexed with (s)- wiskostatin, a small molecule inhibitor
Structure: Wiskott-aldrich syndrome protein. Chain: a. Fragment: core autoinhibited domain (gtpase binding domain is covalently linked to the cofilin homology and acidic regions). Synonym: wasp. Engineered: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Gene: wasp (residues 242-310 and 461-492). Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.
NMR struc: 20 models
Authors: J.R.Peterson,L.C.Bickford,D.Morgan,A.S.Kim,O.Ouerfelli, M.W.Kirschner,M.K.Rosen
Key ref:
J.R.Peterson et al. (2004). Chemical inhibition of N-WASP by stabilization of a native autoinhibited conformation. Nat Struct Mol Biol, 11, 747-755. PubMed id: 15235593 DOI: 10.1038/nsmb796
Date:
11-May-04     Release date:   13-Jul-04    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
P42768  (WASP_HUMAN) -  Wiskott-Aldrich syndrome protein
Seq:
Struc:
502 a.a.
107 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 35 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     positive regulation of Arp2/3 complex-mediated actin nucleation   2 terms 
  Biochemical function     actin binding     1 term  

 

 
DOI no: 10.1038/nsmb796 Nat Struct Mol Biol 11:747-755 (2004)
PubMed id: 15235593  
 
 
Chemical inhibition of N-WASP by stabilization of a native autoinhibited conformation.
J.R.Peterson, L.C.Bickford, D.Morgan, A.S.Kim, O.Ouerfelli, M.W.Kirschner, M.K.Rosen.
 
  ABSTRACT  
 
Current drug discovery efforts focus primarily on proteins with defined enzymatic or small molecule binding sites. Autoregulatory domains represent attractive alternative targets for small molecule inhibitors because they also occur in noncatalytic proteins and because allosteric inhibitors may avoid specificity problems inherent in active site-directed inhibitors. We report here the identification of wiskostatin, a chemical inhibitor of the neural Wiskott-Aldrich syndrome protein (N-WASP). Wiskostatin interacts with a cleft in the regulatory GTPase-binding domain (GBD) of WASP in the solution structure of the complex. Wiskostatin induces folding of the isolated, unstructured GBD into its autoinhibited conformation, suggesting that wiskostatin functions by stabilizing N-WASP in its autoinhibited state. The use of small molecules to bias conformational equilibria represents a potentially general strategy for chemical inhibition of autoinhibited proteins, even in cases where such sites have not been naturally evolved in a target.
 
  Selected figure(s)  
 
Figure 1.
Figure 1. Structure and regulation of the Wiskott-Aldrich syndrome protein family. (a) Domain structure of N-WASP, WASP and WASP constructs used in this study. The following functional domains are indicated: WH1 (WASP-homology 1) or EVH1 (Ena, vasodilator-stimulated phosphoprotein, or VASP-homology 1); a highly basic region (BR); the GTPase-binding domain (GBD); a proline-rich region; and a C-terminal VCA region (verprolin homology, central hydrophobic region, acidic region). WASP and N-WASP share 70% sequence identity over the residues included in mini-WASP. (b) Activation of WASP proteins by Cdc42 and PIP[2] requires the dissociation of an intramolecular autoinhibitory interaction between the GBD and VCA elements, allowing the VCA element to activate Arp2/3 complex.
Figure 2.
Figure 2. Structure and potency of wiskostatin and derivatives. (a) Dose-dependent inhibition of actin polymerization in cytoplasmic extract by (S)-wiskostatin. The increase in pyrene-actin fluorescence in response to PIP[2] addition reflects polymerization of pyrene-actin into filaments. The maximal rate of polymerization is inhibited by 50% in the presence of 5 M (S)-wiskostatin and almost completely by 10 M (S)-wiskostatin with respect to vehicle control (DMSO). Additional concentrations tested are not shown. (b) For each compound, the concentration required to inhibit the actin polymerization rate in PIP[2]-stimulated extract to 50% of the maximum control rate is shown. Atoms distinguishing control compounds from wiskostatin are in large print.
 
  The above figures are reprinted by permission from Macmillan Publishers Ltd: Nat Struct Mol Biol (2004, 11, 747-755) copyright 2004.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
21326322 A.Nürnberg, T.Kitzing, and R.Grosse (2011).
Nucleating actin for invasion.
  Nat Rev Cancer, 11, 177-187.  
21383498 J.S.Orange, S.Roy-Ghanta, E.M.Mace, S.Maru, G.D.Rak, K.B.Sanborn, A.Fasth, R.Saltzman, A.Paisley, L.Monaco-Shawver, P.P.Banerjee, and R.Pandey (2011).
IL-2 induces a WAVE2-dependent pathway for actin reorganization that enables WASp-independent human NK cell function.
  J Clin Invest, 121, 1535-1548.  
21262381 R.van der Meel, M.H.Symons, R.Kudernatsch, R.J.Kok, R.M.Schiffelers, G.Storm, W.M.Gallagher, and A.T.Byrne (2011).
The VEGF/Rho GTPase signalling pathway: a promising target for anti-angiogenic/anti-invasion therapy.
  Drug Discov Today, 16, 219-228.  
21427700 S.J.King, D.C.Worth, T.M.Scales, J.Monypenny, G.E.Jones, and M.Parsons (2011).
β1 integrins regulate fibroblast chemotaxis through control of N-WASP stability.
  EMBO J, 30, 1705-1718.  
  20585499 A.Dovas, and D.Cox (2010).
Regulation of WASp by phosphorylation: Activation or other functions?
  Commun Integr Biol, 3, 101-105.  
20808845 D.Vingadassalom, K.G.Campellone, M.J.Brady, B.Skehan, S.E.Battle, D.Robbins, A.Kapoor, G.Hecht, S.B.Snapper, and J.M.Leong (2010).
Enterohemorrhagic E. coli requires N-WASP for efficient type III translocation but not for EspFU-mediated actin pedestal formation.
  PLoS Pathog, 6, 0.  
20130240 H.Stabile, C.Carlino, C.Mazza, S.Giliani, S.Morrone, L.D.Notarangelo, L.D.Notarangelo, A.Santoni, and A.Gismondi (2010).
Impaired NK-cell migration in WAS/XLT patients: role of Cdc42/WASp pathway in the control of chemokine-induced beta2 integrin high-affinity state.
  Blood, 115, 2818-2826.  
20534520 P.P.Lie, A.Y.Chan, D.D.Mruk, W.M.Lee, and C.Y.Cheng (2010).
Restricted Arp3 expression in the testis prevents blood-testis barrier disruption during junction restructuring at spermatogenesis.
  Proc Natl Acad Sci U S A, 107, 11411-11416.  
20403871 P.P.Lie, D.D.Mruk, W.M.Lee, and C.Y.Cheng (2010).
Cytoskeletal dynamics and spermatogenesis.
  Philos Trans R Soc Lond B Biol Sci, 365, 1581-1592.  
20714349 S.A.Johnston, and R.C.May (2010).
The human fungal pathogen Cryptococcus neoformans escapes macrophages by a phagosome emptying mechanism that is inhibited by Arp2/3 complex-mediated actin polymerisation.
  PLoS Pathog, 6, 0.  
20533885 S.B.Padrick, and M.K.Rosen (2010).
Physical mechanisms of signal integration by WASP family proteins.
  Annu Rev Biochem, 79, 707-735.  
21070974 T.I.Strochlic, J.Viaud, U.E.Rennefahrt, T.Anastassiadis, and J.R.Peterson (2010).
Phosphoinositides are essential coactivators for p21-activated kinase 1.
  Mol Cell, 40, 493-500.  
19961849 T.Inoue, P.P.Pattabiraman, D.L.Epstein, and P.Vasantha Rao (2010).
Effects of chemical inhibition of N-WASP, a critical regulator of actin polymerization on aqueous humor outflow through the conventional pathway.
  Exp Eye Res, 90, 360-367.  
19808890 A.Dovas, J.C.Gevrey, A.Grossi, H.Park, W.Abou-Kheir, and D.Cox (2009).
Regulation of podosome dynamics by WASp phosphorylation: implication in matrix degradation and chemotaxis in macrophages.
  J Cell Sci, 122, 3873-3882.  
18704644 C.C.Calhoun, Y.C.Lu, J.Song, and R.Chiu (2009).
Knockdown endogenous CypA with siRNA in U2OS cells results in disruption of F-actin structure and alters tumor phenotype.
  Mol Cell Biochem, 320, 35-43.  
19741094 H.Park, and D.Cox (2009).
Cdc42 regulates Fc gamma receptor-mediated phagocytosis through the activation and phosphorylation of Wiskott-Aldrich syndrome protein (WASP) and neural-WASP.
  Mol Biol Cell, 20, 4500-4508.  
19759398 H.Yamada, S.Padilla-Parra, S.J.Park, T.Itoh, M.Chaineau, I.Monaldi, O.Cremona, F.Benfenati, P.De Camilli, M.Coppey-Moisan, M.Tramier, T.Galli, and K.Takei (2009).
Dynamic interaction of amphiphysin with N-WASP regulates actin assembly.
  J Biol Chem, 284, 34244-34256.  
18842965 J.C.Porter, and A.Hall (2009).
Epithelial ICAM-1 and ICAM-2 regulate the egression of human T cells across the bronchial epithelium.
  FASEB J, 23, 492-502.  
19561083 M.Cammer, J.C.Gevrey, M.Lorenz, A.Dovas, J.Condeelis, and D.Cox (2009).
The mechanism of CSF-1-induced Wiskott-Aldrich syndrome protein activation in vivo: a role for phosphatidylinositol 3-kinase and Cdc42.
  J Biol Chem, 284, 23302-23311.  
19593382 W.Greene, and S.J.Gao (2009).
Actin dynamics regulate multiple endosomal steps during Kaposi's sarcoma-associated herpesvirus entry and trafficking in endothelial cells.
  PLoS Pathog, 5, e1000512.  
18430734 A.M.Wegner, C.A.Nebhan, L.Hu, D.Majumdar, K.M.Meier, A.M.Weaver, and D.J.Webb (2008).
N-wasp and the arp2/3 complex are critical regulators of actin in the development of dendritic spines and synapses.
  J Biol Chem, 283, 15912-15920.  
18667055 G.Bompard, G.Rabeharivelo, and N.Morin (2008).
Inhibition of cytokinesis by wiskostatin does not rely on N-WASP/Arp2/3 complex pathway.
  BMC Cell Biol, 9, 42.  
18374167 J.C.Porter (2008).
Epithelial Rho GTPases and the transepithelial migration of lymphocytes.
  Methods Enzymol, 439, 205-217.  
18995840 S.B.Padrick, H.C.Cheng, A.M.Ismail, S.C.Panchal, L.K.Doolittle, S.Kim, B.M.Skehan, J.Umetani, C.A.Brautigam, J.M.Leong, and M.K.Rosen (2008).
Hierarchical regulation of WASP/WAVE proteins.
  Mol Cell, 32, 426-438.  
17405146 C.Bacon, V.Lakics, L.Machesky, and M.Rumsby (2007).
N-WASP regulates extension of filopodia and processes by oligodendrocyte progenitors, oligodendrocytes, and Schwann cells-implications for axon ensheathment at myelination.
  Glia, 55, 844-858.  
17157450 D.R.Brown, and L.D.Price (2007).
Characterization of Salmonella enterica serovar Typhimurium DT104 invasion in an epithelial cell line (IPEC J2) from porcine small intestine.
  Vet Microbiol, 120, 328-333.  
17182853 J.A.Legg, G.Bompard, J.Dawson, H.L.Morris, N.Andrew, L.Cooper, S.A.Johnston, G.Tramountanis, and L.M.Machesky (2007).
N-WASP involvement in dorsal ruffle formation in mouse embryonic fibroblasts.
  Mol Biol Cell, 18, 678-687.  
17942688 J.Heuvingh, M.Franco, P.Chavrier, and C.Sykes (2007).
ARF1-mediated actin polymerization produces movement of artificial vesicles.
  Proc Natl Acad Sci U S A, 104, 16928-16933.  
17592133 K.Hybiske, and R.S.Stephens (2007).
Mechanisms of host cell exit by the intracellular bacterium Chlamydia.
  Proc Natl Acad Sci U S A, 104, 11430-11435.  
17084917 R.Ganeshan, K.Nowotarski, A.Di, D.J.Nelson, and K.L.Kirk (2007).
CFTR surface expression and chloride currents are decreased by inhibitors of N-WASP and actin polymerization.
  Biochim Biophys Acta, 1773, 192-200.  
16687574 A.Robert, N.Smadja-Lamère, M.C.Landry, C.Champagne, R.Petrie, N.Lamarche-Vane, H.Hosoya, and J.N.Lavoie (2006).
Adenovirus E4orf4 hijacks rho GTPase-dependent actin dynamics to kill cells: a role for endosome-associated actin assembly.
  Mol Biol Cell, 17, 3329-3344.  
16687395 C.Haller, S.Rauch, N.Michel, S.Hannemann, M.J.Lehmann, O.T.Keppler, and O.T.Fackler (2006).
The HIV-1 pathogenicity factor Nef interferes with maturation of stimulatory T-lymphocyte contacts by modulation of N-Wasp activity.
  J Biol Chem, 281, 19618-19630.  
16632257 J.R.Peterson, A.M.Lebensohn, H.E.Pelish, and M.W.Kirschner (2006).
Biochemical suppression of small-molecule inhibitors: a strategy to identify inhibitor targets and signaling pathway components.
  Chem Biol, 13, 443-452.  
16894562 R.Schreck, and U.R.Rapp (2006).
Raf kinases: oncogenesis and drug discovery.
  Int J Cancer, 119, 2261-2271.  
15800060 A.I.Ivanov, D.Hunt, M.Utech, A.Nusrat, and C.A.Parkos (2005).
Differential roles for actin polymerization and a myosin II motor in assembly of the epithelial apical junctional complex.
  Mol Biol Cell, 16, 2636-2650.  
15821030 D.W.Leung, and M.K.Rosen (2005).
The nucleotide switch in Cdc42 modulates coupling between the GTPase-binding and allosteric equilibria of Wiskott-Aldrich syndrome protein.
  Proc Natl Acad Sci U S A, 102, 5685-5690.  
16246732 L.Hemsath, R.Dvorsky, D.Fiegen, M.F.Carlier, and M.R.Ahmadian (2005).
An electrostatic steering mechanism of Cdc42 recognition by Wiskott-Aldrich syndrome proteins.
  Mol Cell, 20, 313-324.
PDB code: 2atx
16006560 M.A.Chellaiah (2005).
Regulation of actin ring formation by rho GTPases in osteoclasts.
  J Biol Chem, 280, 32930-32943.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB code is shown on the right.