PDBsum entry 1rk4

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protein Protein-protein interface(s) links
Ion transport/membrane protein PDB id
Protein chains
213 a.a. *
Waters ×342
* Residue conservation analysis
PDB id:
Name: Ion transport/membrane protein
Title: Crystal structure of a soluble dimeric form of oxidised clic1
Structure: Chloride intracellular channel protein 1. Chain: a, b. Synonym: nuclear chloride ion channel 27. Ncc27. P64 clcp. Chloride channel abp. P64clcp. Chloride channel abp. Engineered: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Gene: clic1, ncc27. Expressed in: escherichia coli. Expression_system_taxid: 562.
Biol. unit: Dimer (from PQS)
1.79Å     R-factor:   0.178     R-free:   0.213
Authors: D.R.Littler,S.J.Harrop,W.D.Fairlie,L.J.Brown,G.J.Pankhurst, S.Pankhurst,M.Z.Demaere,T.J.Campbell,A.R.Bauskin,R.Tonini, M.Mazzanti,S.N.Breit,P.M.Curmi
Key ref:
D.R.Littler et al. (2004). The intracellular chloride ion channel protein CLIC1 undergoes a redox-controlled structural transition. J Biol Chem, 279, 9298-9305. PubMed id: 14613939 DOI: 10.1074/jbc.M308444200
20-Nov-03     Release date:   02-Dec-03    
Go to PROCHECK summary

Protein chains
Pfam   ArchSchema ?
O00299  (CLIC1_HUMAN) -  Chloride intracellular channel protein 1
241 a.a.
213 a.a.
Key:    PfamA domain  Secondary structure  CATH domain

 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     blood microparticle   14 terms 
  Biological process     transport   9 terms 
  Biochemical function     protein binding     4 terms  


DOI no: 10.1074/jbc.M308444200 J Biol Chem 279:9298-9305 (2004)
PubMed id: 14613939  
The intracellular chloride ion channel protein CLIC1 undergoes a redox-controlled structural transition.
D.R.Littler, S.J.Harrop, W.D.Fairlie, L.J.Brown, G.J.Pankhurst, S.Pankhurst, M.Z.DeMaere, T.J.Campbell, A.R.Bauskin, R.Tonini, M.Mazzanti, S.N.Breit, P.M.Curmi.
Most proteins adopt a well defined three-dimensional structure; however, it is increasingly recognized that some proteins can exist with at least two stable conformations. Recently, a class of intracellular chloride ion channel proteins (CLICs) has been shown to exist in both soluble and integral membrane forms. The structure of the soluble form of CLIC1 is typical of a soluble glutathione S-transferase superfamily protein but contains a glutaredoxin-like active site. In this study we show that on oxidation CLIC1 undergoes a reversible transition from a monomeric to a non-covalent dimeric state due to the formation of an intramolecular disulfide bond (Cys-24-Cys-59). We have determined the crystal structure of this oxidized state and show that a major structural transition has occurred, exposing a large hydrophobic surface, which forms the dimer interface. The oxidized CLIC1 dimer maintains its ability to form chloride ion channels in artificial bilayers and vesicles, whereas a reducing environment prevents the formation of ion channels by CLIC1. Mutational studies show that both Cys-24 and Cys-59 are required for channel activity.
  Selected figure(s)  
Figure 2.
FIG. 2. Structure of the oxidized CLIC1 dimer. A, stereo backbone of the CLIC1 dimer; green, A subunit; red, B subunit. In the A subunit every 10th residue is labeled. B, electron density of the intramolecular disulfide bond between Cys-24 and Cys-59 contoured at 1 . The CLIC1 dimer is viewed along (C) and perpendicular (D) to the pseudo 2-fold axis; are shown helices, A subunit (red) and B subunit (green), and intramolecular disulfide bonds (yellow). E, ClustalW (24) alignment of the CLIC family. The secondary structure is shown for both monomeric (red, helices; yellow, -strands) and dimeric (blue, helices) forms. Conserved regions are shaded; green, putative transmembrane regions; yellow, Cys; cream, Gly. Features unique to CLIC1 are in blue. Ramachandran distances (see "Experimental Procedures") for the monomer to dimer transition are plotted above its sequence. Figures were made with SETOR (25), MOLSCRIPT (26), RASTER3D (27), and CONSCRIPT (28).
Figure 3.
FIG. 3. Structural transition of CLIC1 between the monomeric and the dimeric forms. Representations of reduced monomeric form of CLIC1 (A) and a subunit of the oxidized dimeric form (B). C, backbone superposition of CLIC1 for the reduced monomeric (green) and the oxidized dimeric (magenta) states. Ramachandran distances for residues 23-234 are mapped onto the backbone of the monomeric (D) and dimeric (E) forms. The color gradient, from gray to pink represents Ramachandran distances from 0° to 180°. Residues not observed in the dimer are colored gold. F, Ramachandran plot of residues within the N-domain with Ramachandran distances greater than 35^0 between the two structures. Monomer - co-ordinates are plotted as orange squares, and dimer co-ordinates are in black with a connecting line. The figures were made with SETOR (25), MOLSCRIPT (26), RASTER3D (27), and GRASP (29).
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2004, 279, 9298-9305) copyright 2004.  
  Figures were selected by the author.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
  21323602 A.F.Dulhunty, R.Hewawasam, D.Liu, M.G.Casarotto, and P.G.Board (2011).
Regulation of the cardiac muscle ryanodine receptor by glutathione transferases.
  Drug Metab Rev, 43, 236-252.  
19850480 F.Sesti, S.Liu, and S.Q.Cai (2010).
Oxidation of potassium channels by ROS: a general mechanism of aging and neurodegeneration?
  Trends Cell Biol, 20, 45-51.  
  21040583 J.J.Tung, and J.Kitajewski (2010).
Chloride intracellular channel 1 functions in endothelial cell growth and migration.
  J Angiogenes Res, 2, 23.  
19634988 M.A.Wouters, S.W.Fan, and N.L.Haworth (2010).
Disulfides as redox switches: from molecular mechanisms to functional significance.
  Antioxid Redox Signal, 12, 53-91.  
20591649 P.N.Bryan, and J.Orban (2010).
Proteins that switch folds.
  Curr Opin Struct Biol, 20, 482-488.  
  20824052 R.Brauer, L.C.Wang, S.T.Woon, D.J.Bridewell, K.Henare, D.Malinger, B.D.Palmer, S.N.Vogel, C.Kieda, S.M.Tijono, and L.M.Ching (2010).
Labeling of oxidizable proteins with a photoactivatable analog of the antitumor agent DMXAA: evidence for redox signaling in its mode of action.
  Neoplasia, 12, 755-765.  
19448624 A.Shukla, M.Malik, C.Cataisson, Y.Ho, T.Friesen, K.S.Suh, and S.H.Yuspa (2009).
TGF-beta signalling is regulated by Schnurri-2-dependent nuclear translocation of CLIC4 and consequent stabilization of phospho-Smad2 and 3.
  Nat Cell Biol, 11, 777-784.  
19776349 B.Ponsioen, L.van Zeijl, M.Langeslag, M.Berryman, D.Littler, K.Jalink, and W.H.Moolenaar (2009).
Spatiotemporal regulation of chloride intracellular channel protein CLIC4 by RhoA.
  Mol Biol Cell, 20, 4664-4672.  
19650640 S.H.Stoychev, C.Nathaniel, S.Fanucchi, M.Brock, S.Li, K.Asmus, V.L.Woods, and H.W.Dirr (2009).
Structural dynamics of soluble chloride intracellular channel protein CLIC1 examined by amide hydrogen-deuterium exchange mass spectrometry.
  Biochemistry, 48, 8413-8421.  
19598234 S.W.Fan, R.A.George, N.L.Haworth, L.L.Feng, J.Y.Liu, and M.A.Wouters (2009).
Conformational changes in redox pairs of protein structures.
  Protein Sci, 18, 1745-1765.  
19356589 X.Meng, G.Wang, C.Viero, Q.Wang, W.Mi, X.D.Su, T.Wagenknecht, A.J.Williams, Z.Liu, and C.C.Yin (2009).
CLIC2-RyR1 interaction and structural characterization by cryo-electron microscopy.
  J Mol Biol, 387, 320-334.  
18175373 A.Rahman, S.G.Kumar, S.W.Kim, H.J.Hwang, Y.M.Baek, S.H.Lee, H.S.Hwang, Y.H.Shon, K.S.Nam, and J.W.Yun (2008).
Proteomic analysis for inhibitory effect of chitosan oligosaccharides on 3T3-L1 adipocyte differentiation.
  Proteomics, 8, 569-581.  
17985355 D.R.Littler, S.J.Harrop, L.J.Brown, G.J.Pankhurst, A.V.Mynott, P.Luciani, R.A.Mandyam, M.Mazzanti, S.Tanda, M.A.Berryman, S.N.Breit, and P.M.Curmi (2008).
Comparison of vertebrate and invertebrate CLIC proteins: the crystal structures of Caenorhabditis elegans EXC-4 and Drosophila melanogaster DmCLIC.
  Proteins, 71, 364-378.
PDB codes: 2yv7 2yv9
19111067 R.R.Thangudu, M.Manoharan, N.Srinivasan, F.Cadet, R.Sowdhamini, and B.Offmann (2008).
Analysis on conservation of disulphide bonds and their structural features in homologous protein domain families.
  BMC Struct Biol, 8, 55.  
  18007051 B.A.Cromer, M.A.Gorman, G.Hansen, J.J.Adams, M.Coggan, P.G.Board, and M.W.Parker (2007).
Expression, purification, crystallization and preliminary X-ray diffraction analysis of chloride intracellular channel 2 (CLIC2).
  Acta Crystallogr Sect F Struct Biol Cryst Commun, 63, 961-963.  
17326840 B.Ulmasov, J.Bruno, P.G.Woost, and J.C.Edwards (2007).
Tissue and subcellular distribution of CLIC1.
  BMC Cell Biol, 8, 8.  
16495342 C.S.Sevier, and C.A.Kaiser (2006).
Disulfide transfer between two conserved cysteine pairs imparts selectivity to protein oxidation by Ero1.
  Mol Biol Cell, 17, 2256-2266.  
16339885 H.Singh, and R.H.Ashley (2006).
Redox regulation of CLIC1 by cysteine residues associated with the putative channel pore.
  Biophys J, 90, 1628-1638.  
17347778 J.C.Edwards (2006).
The CLIC1 chloride channel is regulated by the cystic fibrosis transmembrane conductance regulator when expressed in Xenopus oocytes.
  J Membr Biol, 213, 39-46.  
16176272 D.R.Littler, N.N.Assaad, S.J.Harrop, L.J.Brown, G.J.Pankhurst, P.Luciani, M.I.Aguilar, M.Mazzanti, M.A.Berryman, S.N.Breit, and P.M.Curmi (2005).
Crystal structure of the soluble form of the redox-regulated chloride ion channel protein CLIC4.
  FEBS J, 272, 4996-5007.
PDB code: 2ahe
15822171 J.D.Hayes, J.U.Flanagan, and I.R.Jowsey (2005).
Glutathione transferases.
  Annu Rev Pharmacol Toxicol, 45, 51-88.  
15501681 N.J.Marianayagam, M.Sunde, and J.M.Matthews (2004).
The power of two: protein dimerization in biology.
  Trends Biochem Sci, 29, 618-625.  
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 codes are shown on the right.