PDBsum entry 1qvy

Signaling protein inhibitor

PDB id: 1qvy

Contents

Protein chains

138 a.a. *

Ligands

SO4 ×8

Waters ×779

* Residue conservation analysis

PDB id:

1qvy

Name:

Signaling protein inhibitor

Title:

Crystal structure of rhogdi k(199,200)r double mutant

Structure:

Rho gdp-dissociation inhibitor 1. Chain: a, b, c, d. Fragment: c-terminal domain. Synonym: rho gdi 1. Rho-gdi alpha. Rho gdp dissociation inhibitor. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.60Å R-factor: 0.173 R-free: 0.210

Authors:

J.Czepas,Y.Devedjiev,D.Krowarsh,U.Derewenda,Z.S.Derewenda

Key ref:

J.Czepas et al. (2004). The impact of Lys-->Arg surface mutations on the crystallization of the globular domain of RhoGDI. Acta Crystallogr D Biol Crystallogr, 60, 275-280. PubMed id: 14747703 DOI: 10.1107/S0907444903026271

Date:

29-Aug-03 Release date: 10-Feb-04

Protein chains

P52565  (GDIR1_HUMAN) -  Rho GDP-dissociation inhibitor 1 from Homo sapiens

Seq: 204 a.a.

Struc: 138 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

DOI no: 10.1107/S0907444903026271

Acta Crystallogr D Biol Crystallogr 60:275-280 (2004)

PubMed id: 14747703

The impact of Lys-->Arg surface mutations on the crystallization of the globular domain of RhoGDI.

J.Czepas, Y.Devedjiev, D.Krowarsch, U.Derewenda, J.Otlewski, Z.S.Derewenda.

ABSTRACT

The potential of rational surface mutagenesis for enhanced protein crystallization is being probed in an ongoing effort. In previous work, it was hypothesized that residues with high conformational entropy such as Glu and Lys are suitable targets for surface mutagenesis, as they are rarely incorporated in crystal contacts or protein-protein interfaces. Previous experiments using Lys-->Ala, Glu-->Ala and Glu-->Asp mutants confirmed that mutated proteins were more likely to crystallize. In the present paper, the usefulness of Lys-->Arg mutations is studied. Several mutations of the globular domain of human RhoGDI were generated, including the single mutants K105R, K113R, K127R, K138R and K141R, the double mutants K(98,99)R and K(199,200)R and the triple mutants K(98,99,105)R and K(135,138,141)R. It is shown that Lys-->Arg mutants are more likely to crystallize than the wild-type protein, although not as likely as Lys-->Ala mutants. Out of the nine mutants tested, five produced diffracting crystals, including the K(199,200)R double mutant, which crystallized in a new space group and exceeded by approximately 1.0 A the resolution of the diffraction of the wild-type crystal. Major crystal contacts in the new lattice were created by the mutated epitope.

Selected figure(s)

Figure 3.
Figure 3 Electron-density map in the region of the lattice contact contoured at the 1 level, involving mutated Arg199, its symmetry-related pair Arg199 sym as well as the charge constellation including the negatively charged residues Asp204, Asp204 sym and two solvent sulfates. Additional hydrogen bonds involving Arg200, Ser174 and their symmetry-related partners contribute to stability of the lattice contact. The figure was prepared with BOBSCRIPT (Esnouf, 1997[Esnouf, R. M. (1997). J. Mol. Graph. 15, 132-143.]).

The above figure is reprinted by permission from the IUCr: Acta Crystallogr D Biol Crystallogr (2004, 60, 275-280) copyright 2004.

Figure was selected by the author.