spacer
spacer

PDBsum entry 1rrp

Go to PDB code: 
Top Page protein ligands metals Protein-protein interface(s) links
Complex (small gtpase/nuclear protein) PDB id
1rrp
Jmol
Contents
Protein chains
204 a.a. *
134 a.a. *
180 a.a. *
Ligands
GNP ×2
Metals
_MG ×2
Waters ×36
* Residue conservation analysis

References listed in PDB file
Key reference
Title Structure of a ran-Binding domain complexed with ran bound to a gtp analogue: implications for nuclear transport.
Authors I.R.Vetter, C.Nowak, T.Nishimoto, J.Kuhlmann, A.Wittinghofer.
Ref. Nature, 1999, 398, 39-46. [DOI no: 10.1038/17969]
PubMed id 10078529
Abstract
The protein Ran is a small GTP-binding protein that binds to two types of effector inside the cell: Ran-binding proteins, which have a role in terminating export processes from the nucleus to the cytoplasm, and importin-beta-like molecules that bind cargo proteins during nuclear transport. The Ran-binding domain is a conserved sequence motif found in several proteins that participate in these transport processes. The Ran-binding protein RanBP2 contains four of these domains and constitutes a large part of the cytoplasmic fibrils that extend from the nuclear-pore complex. The structure of Ran bound to a non-hydrolysable GTP analogue (Ran x GppNHp) in complex with the first Ran-binding domain (RanBD1) of human RanBP2 reveals not only that RanBD1 has a pleckstrin-homology domain fold, but also that the switch-I region of Ran x GppNHp resembles the canonical Ras GppNHp structure and that the carboxy terminus of Ran is wrapped around RanBD1, contacting a basic patch on RanBD1 through its acidic end. This molecular 'embrace' enables RanBDs to sequester the Ran carboxy terminus, triggering the dissociation of Ran x GTP from importin-beta-related transport factors and facilitating GTP hydrolysis by the GTPase-activating protein ranGAP. Such a mechanism represents a new type of switch mechanism and regulatory protein-protein interaction for a Ras-related protein.
Figure 1.
Figure 1: Representative electron density around switch I in Ran dot-GppNHp and the conserved WKER motif of RanBD1 (residues 57–60). Residues from RanBD1 are represented by white carbon traces, residues from Ran by yellow carbon traces. The omit map is contoured at 1.1 . The figure was prepared with BOBSCRIPT^46.
Figure 5.
Figure 5: Molecular embrace and the DEDDDL motif. Surface representation of RanBD1, showing the basic region where the DEDDDL motif of Ran is expected to bind after the C terminus wraps itself around RanBD1. Ran is shown as a backbone (green), and GppNHp and the magnesium ion as ball and stick. The figure was produced using GRASP^50.
The above figures are reprinted by permission from Macmillan Publishers Ltd: Nature (1999, 398, 39-46) copyright 1999.
Secondary reference #1
Title Ranbp1 is crucial for the release of rangtp from importin beta-Related nuclear transport factors.
Authors F.R.Bischoff, D.Görlich.
Ref. FEBS Lett, 1997, 419, 249-254. [DOI no: 10.1016/S0014-5793(97)01467-1]
PubMed id 9428644
Full text Abstract
Figure 3.
Fig. 3. A: RanBP1 directly induces disassembly of the transportin·RanGTP and the importin α·CAS·RanGTP complexes. Transportin, tagged with the IgG binding z-domain from protein A, was bound to IgG-nitrocellulose (for details of the assay see Section 2). Then, Ran[γ-^32P]GTP was allowed for 15 min to bind the immobilised transportin and non-bound radioactivity was washed away. Then, excess of non-immobilised transportin (i.e. lacking the z-tag) was added as a trap for RanGTP released from immobilised transportin. Then buffer or 100 nM RanBP1 was added and after the indicated time intervals, the radioactivity was determined on the filter and in the supernatant. Dissociation of RanGTP from transportin is given as the ratio between filter-bound and free radioactivity. B: CAS was expressed in E. coli, a lysate was prepared and 0.5 μM importin α was added. Three ml of the mixture (corresponding to 30 ml bacterial culture) was bound to z-tagged RanGTP that had been immobilised to 30 μl IgG Sepharose. The Sepharose was washed 5 times with 1 ml low salt binding buffer (containing 0.1 M NaCl) and split into three. Elution was performed with 80 μl of either high salt buffer (1 M NaCl), low salt buffer (0.1 M salt), or with low salt buffer plus 1 nmole RanBP1. The figure shows the Coomassie stained gel loaded with 2 μl of the lysate and 1/10 of the eluates. Note that CAS and importin α could be eluted from RanGTP in the absence of RanBP1 only at high salt concentration. In contrast, in the presence of RanBP1 this disassembly also occurred at physiological ionic strength. z-tagged Ran remained bound to the IgG Sepharose at low or high salt.
Figure 4.
Fig. 4. Model for the disassembly of transport factor·RanGTP complexes. Disassembly of complexes of Ran·GTP and A: CAS; B: transportin; and C: importin-β. For details see main text.
The above figures are reproduced from the cited reference with permission from the Federation of European Biochemical Societies
Secondary reference #2
Title Dynamic and equilibrium studies on the interaction of ran with its effector, Ranbp1.
Authors J.Kuhlmann, I.Macara, A.Wittinghofer.
Ref. Biochemistry, 1997, 36, 12027-12035. [DOI no: 10.1021/bi970524k]
PubMed id 9315840
Full text Abstract
Secondary reference #3
Title A ran-Binding motif in nuclear pore proteins.
Authors E.Hartmann, D.Görlich.
Ref. Trends Cell Biol, 1995, 5, 192-193.
PubMed id 14731447
Abstract
Secondary reference #4
Title A giant nucleopore protein that binds ran/tc4.
Authors N.Yokoyama, N.Hayashi, T.Seki, N.Panté, T.Ohba, K.Nishii, K.Kuma, T.Hayashida, T.Miyata, U.Aebi.
Ref. Nature, 1995, 376, 184-188.
PubMed id 7603572
Abstract
Secondary reference #5
Title The c terminus of the nuclear ran/tc4 gtpase stabilizes the gdp-Bound state and mediates interactions with rcc1, Ran-Gap, And htf9a/ranbp1.
Authors S.A.Richards, K.M.Lounsbury, I.G.Macara.
Ref. J Biol Chem, 1995, 270, 14405-14411.
PubMed id 7782302
Abstract
Secondary reference #6
Title Co-Activation of rangtpase and inhibition of gtp dissociation by ran-Gtp binding protein ranbp1.
Authors F.R.Bischoff, H.Krebber, E.Smirnova, W.Dong, H.Ponstingl.
Ref. EMBO J, 1995, 14, 705-715.
PubMed id 7882974
Abstract
PROCHECK
Go to PROCHECK summary
 Headers