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PDBsum entry 1bg5

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protein links
Ankyrin binding PDB id
1bg5
Jmol
Contents
Protein chain
254 a.a. *
Waters ×147
* Residue conservation analysis
PDB id:
1bg5
Name: Ankyrin binding
Title: Crystal structure of the ankyrin binding domain of alpha-na, k-atpase as a fusion protein with glutathione s-transferase
Structure: Fusion protein of alpha-na,k-atpase with glutathione s-transferase. Chain: a. Fragment: residues 1-218 from glutathione s-transferase, residues 219-254 from minimal alpha-na,k-atpase ankyrin binding domain. Synonym: mab. Engineered: yes
Source: Fragment: residues 219-254. Rattus norvegicus. Norway rat. Organism_taxid: 10116. Organ: blood. Cellular_location: plasma membrane. Expressed in: escherichia coli. Expression_system_taxid: 562.
Biol. unit: Dimer (from PQS)
Resolution:
2.60Å     R-factor:   0.193     R-free:   0.359
Authors: Z.Zhang,P.Devarajan,J.S.Morrow
Key ref:
Z.Zhang et al. (1998). Structure of the ankyrin-binding domain of alpha-Na,K-ATPase. J Biol Chem, 273, 18681-18684. PubMed id: 9668035 DOI: 10.1074/jbc.273.30.18681
Date:
05-Jun-98     Release date:   13-Jan-99    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
P08515  (GST26_SCHJA) -  Glutathione S-transferase class-mu 26 kDa isozyme
Seq:
Struc:
218 a.a.
254 a.a.
Key:    PfamA domain  Secondary structure  CATH domain

 Enzyme reactions 
   Enzyme class: E.C.2.5.1.18  - Glutathione transferase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: RX + glutathione = HX + R-S-glutathione
RX
+ glutathione
= HX
+ R-S-glutathione
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     metabolic process   1 term 
  Biochemical function     transferase activity     2 terms  

 

 
    reference    
 
 
DOI no: 10.1074/jbc.273.30.18681 J Biol Chem 273:18681-18684 (1998)
PubMed id: 9668035  
 
 
Structure of the ankyrin-binding domain of alpha-Na,K-ATPase.
Z.Zhang, P.Devarajan, A.L.Dorfman, J.S.Morrow.
 
  ABSTRACT  
 
The ankyrin 33-residue repeating motif, an L-shaped structure with protruding beta-hairpin tips, mediates specific macromolecular interactions with cytoskeletal, membrane, and regulatory proteins. The association between ankyrin and alpha-Na,K-ATPase, a ubiquitous membrane protein critical to vectorial transport of ions and nutrients, is required to assemble and stabilize Na,K-ATPase at the plasma membrane. alpha-Na,K-ATPase binds both red cell ankyrin (AnkR, a product of the ANK1 gene) and Madin-Darby canine kidney cell ankyrin (AnkG, a product of the ANK3 gene) utilizing residues 142-166 (SYYQEAKSSKIMESFK NMVPQQALV) in its second cytoplasmic domain. Fusion peptides of glutathione S-transferase incorporating these 25 amino acids bind specifically to purified ankyrin (Kd = 118 +/- 50 nM). The three-dimensional structure (2.6 A) of this minimal ankyrin-binding motif, crystallized as the fusion protein, reveals a 7-residue loop with one charged hydrophilic face capping a double beta-strand. Comparison with ankyrin-binding sequences in p53, CD44, neurofascin/L1, and the inositol 1,4,5-trisphosphate receptor suggests that the valency and specificity of ankyrin binding is achieved by the interaction of 5-7-residue surface loops with the beta-hairpin tips of multiple ankyrin repeat units.
 
  Selected figure(s)  
 
Figure 4.
Fig. 4. Three-axis views of the structure of the minimal ankyrin-binding domain of -Na,K-ATPase. The basic structural motif is that of a 7-residue loop on a stalk composed of antiparallel -strands. A, C, and E, surface accessibility depictions showing the amphipathic surfaces formed by the loop. B, D, and F, ribbon diagrams demonstrating the backbone contour. It is envisioned that the 7-residue loop interacts with the -hairpin tips of one or more ankyrin repeat units (see Fig. 5). The arrow in C marks the depth of the structural plane shown in E and F.
Figure 5.
Fig. 5. MAB is an ATPase specific sequence that may interact with one or more ankyrin repeat units. A, sequence alignment of the minimal ankyrin-binding domain (peptide IIA) with all other proteins in GenBankTM and Swiss-Prot. Dashes represent residues identical to human Na,K-ATPase 1 subunit, residues 142-166. This sequence is well conserved among all known Na,K-ATPase subunits and is partially conserved (68% identity) in only one other documented ankyrin-binding protein (gastric H/K-ATPase). There is no homology to the ankyrin-binding sequences in CD44, IP3-R, neurofascin, or p53bp2. However, like MAB, each of the other reported ankyrin-binding sequences are short peptides. B, hypothetical model of how MAB may interact with ankyrin, drawn to scale. Each ankyrin repeat is composed of two -helices and a -hairpin loop (5). Multiple repeat units create a structure in which helix-helix interactions form a central core, whereas the tips of the exposed -hairpin turns provide potential interprotein interaction surfaces. We envision that the 7-residue loop within MAB interacts with site(s) in ankyrin created by the tips of one or more of its -hairpin turns. Because a multiplicity of potential binding pockets would be created by the 13-24 repeat units characteristic of most ankyrins, specific and unique binding sites probably also exist for other short peptide sequences such as those that bestow ankyrin binding activity on other proteins. To effect this binding, we hypothesize that these peptides will assume a loop on a stalk structure similar to that reported here for -Na,K-ATPase.
 
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (1998, 273, 18681-18684) copyright 1998.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
  20512993 P.M.Verhulst, L.M.van der Velden, V.Oorschot, E.E.van Faassen, J.Klumperman, R.H.Houwen, T.G.Pomorski, J.C.Holthuis, and L.W.Klomp (2010).
A flippase-independent function of ATP8B1, the protein affected in familial intrahepatic cholestasis type 1, is required for apical protein expression and microvillus formation in polarized epithelial cells.
  Hepatology, 51, 2049-2060.  
19582774 E.Silva, and P.Soares-da-Silva (2009).
Protein cytoskeleton and overexpression of Na(+),K(+)-ATPase in opossum kidney cells.
  J Cell Physiol, 221, 318-324.  
18283487 Z.Li, and Z.Xie (2009).
The Na/K-ATPase/Src complex and cardiotonic steroid-activated protein kinase cascades.
  Pflugers Arch, 457, 635-644.  
19112491 A.S.Hill, A.Nishino, K.Nakajo, G.Zhang, J.R.Fineman, M.E.Selzer, Y.Okamura, and E.C.Cooper (2008).
Ion channel clustering at the axon initial segment and node of ranvier evolved sequentially in early chordates.
  PLoS Genet, 4, e1000317.  
18780816 L.Corsini, M.Hothorn, K.Scheffzek, M.Sattler, and G.Stier (2008).
Thioredoxin as a fusion tag for carrier-driven crystallization.
  Protein Sci, 17, 2070-2079.  
18599311 M.Doi, and K.Iwasaki (2008).
Na+/K+ ATPase regulates the expression and localization of acetylcholine receptors in a pump activity-independent manner.
  Mol Cell Neurosci, 38, 548-558.  
18768923 P.R.Stabach, P.Devarajan, M.C.Stankewich, S.Bannykh, and J.S.Morrow (2008).
Ankyrin facilitates intracellular trafficking of alpha1-Na+-K+-ATPase in polarized cells.
  Am J Physiol Cell Physiol, 295, C1202-C1214.  
18420589 R.Efendiev, C.E.Budu, A.M.Bertorello, and C.H.Pedemonte (2008).
G-protein-coupled receptor-mediated traffic of Na,K-ATPase to the plasma membrane requires the binding of adaptor protein 1 to a Tyr-255-based sequence in the alpha-subunit.
  J Biol Chem, 283, 17561-17567.  
17575526 M.J.Caplan (2007).
The future of the pump.
  J Clin Gastroenterol, 41, S217-S222.  
17164420 S.M.Paul, M.J.Palladino, and G.J.Beitel (2007).
A pump-independent function of the Na,K-ATPase is required for epithelial junction function and tracheal tube-size control.
  Development, 134, 147-155.  
17060500 A.Das, C.Base, S.Dhulipala, and R.R.Dubreuil (2006).
Spectrin functions upstream of ankyrin in a spectrin cytoskeleton assembly pathway.
  J Cell Biol, 175, 325-335.  
17091212 R.R.Dubreuil (2006).
Functional links between membrane transport and the spectrin cytoskeleton.
  J Membr Biol, 211, 151-161.  
12496082 M.V.Petoukhov, N.A.Eady, K.A.Brown, and D.I.Svergun (2002).
Addition of missing loops and domains to protein models by x-ray solution scattering.
  Biophys J, 83, 3113-3125.  
11276400 A.Aperia (2001).
Regulation of sodium/potassium ATPase activity: impact on salt balance and vascular contractility.
  Curr Hypertens Rep, 3, 165-171.  
11591639 F.C.Serluca, A.Sidow, J.D.Mably, and M.C.Fishman (2001).
Partitioning of tissue expression accompanies multiple duplications of the Na+/K+ ATPase alpha subunit gene.
  Genome Res, 11, 1625-1631.  
11463838 N.Nekrep, M.Geyer, N.Jabrane-Ferrat, and B.M.Peterlin (2001).
Analysis of ankyrin repeats reveals how a single point mutation in RFXANK results in bare lymphocyte syndrome.
  Mol Cell Biol, 21, 5566-5576.  
11163135 V.Bennett, and L.Chen (2001).
Ankyrins and cellular targeting of diverse membrane proteins to physiological sites.
  Curr Opin Cell Biol, 13, 61-67.  
11264588 Y.H.Han, Y.H.Chung, T.Y.Kim, S.J.Hong, J.D.Choi, and Y.J.Chung (2001).
Crystallization of Clonorchis sinensis 26 kDa glutathione S-transferase and its fusion proteins with peptides of different lengths.
  Acta Crystallogr D Biol Crystallogr, 57, 579-581.  
10737894 D.Zhu, and L.Y.Bourguignon (2000).
Interaction between CD44 and the repeat domain of ankyrin promotes hyaluronic acid-mediated ovarian tumor cell migration.
  J Cell Physiol, 183, 182-195.  
10893266 L.Y.Bourguignon, H.Zhu, L.Shao, and Y.W.Chen (2000).
Ankyrin-Tiam1 interaction promotes Rac1 signaling and metastatic breast tumor cell invasion and migration.
  J Cell Biol, 150, 177-191.  
11090820 M.le Maire, P.Champeil, and J.V.Moller (2000).
Interaction of membrane proteins and lipids with solubilizing detergents.
  Biochim Biophys Acta, 1508, 86.  
10791978 R.R.Dubreuil, P.Wang, S.Dahl, J.Lee, and L.S.Goldstein (2000).
Drosophila beta spectrin functions independently of alpha spectrin to polarize the Na,K ATPase in epithelial cells.
  J Cell Biol, 149, 647-656.  
10423269 L.Y.Bourguignon, H.Zhu, L.Shao, D.Zhu, and Y.W.Chen (1999).
Rho-kinase (ROK) promotes CD44v(3,8-10)-ankyrin interaction and tumor cell migration in metastatic breast cancer cells.
  Cell Motil Cytoskeleton, 43, 269-287.  
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.