PDBsum entry: 1htj

Signaling protein

PDB id: 1htj

Contents

Protein chain

182 a.a. *

Waters ×84

* Residue conservation analysis

PDB id:

1htj

Name:

Signaling protein

Title:

Structure of the rgs-like domain from pdz-rhogef

Structure:

Kiaa0380. Chain: f. Fragment: rgs-like domain (residues 281-490). Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: kiaa0380. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.20Å R-factor: 0.223 R-free: 0.258

Authors:

K.L.Longenecker,M.E.Lewis,H.Chikumi,J.S.Gutkind, Z.S.Derewenda

Key ref:

K.L.Longenecker et al. (2001). Structure of the RGS-like domain from PDZ-RhoGEF: linking heterotrimeric g protein-coupled signaling to Rho GTPases. Structure, 9, 559-569. PubMed id: 11470431 DOI: 10.1016/S0969-2126(01)00620-7

Date:

29-Dec-00 Release date: 11-Jul-01

Protein chain

O15085  (ARHGB_HUMAN) -  Rho guanine nucleotide exchange factor 11

Seq: 1522 a.a.

Struc: 182 a.a.*

* PDB and UniProt seqs differ at 3 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 

• Cellular component: cytoplasm (1 term)
• Biological process: termination of G-protein coupled receptor signaling pathway (1 term)
• Biochemical function: Rho guanyl-nucleotide exchange factor activity (1 term)

DOI no: 10.1016/S0969-2126(01)00620-7

Structure 9:559-569 (2001)

PubMed id: 11470431

Structure of the RGS-like domain from PDZ-RhoGEF: linking heterotrimeric g protein-coupled signaling to Rho GTPases.

K.L.Longenecker, M.E.Lewis, H.Chikumi, J.S.Gutkind, Z.S.Derewenda.

ABSTRACT

BACKGROUND: The multidomain PDZ-RhoGEF is one of many known guanine nucleotide exchange factors that upregulate Rho GTPases. PDZ-RhoGEF and related family members play a critical role in a molecular signaling pathway from heterotrimeric G protein-coupled receptors to Rho proteins. A approximately 200 residue RGS-like (RGSL) domain in PDZ-RhoGEF and its homologs is responsible for the direct association with Galpha12/13 proteins. To better understand structure-function relationships, we initiated crystallographic studies of the RGSL domain from human PDZ-RhoGEF. RESULTS: A recombinant construct of the RGSL domain was expressed in Escherichia coli and purified, but it did not crystallize. Alternative constructs were designed based on a novel strategy of targeting lysine and glutamic acid residues for mutagenesis to alanine. A triple-point mutant functionally identical to the wild-type protein was crystallized, and its structure was determined by the MAD method using Se-methionine (Se-Met) incorporation. A molecular model of the RGSL domain was refined at 2.2 A resolution, revealing an all-helical tertiary fold with the mutations located at intermolecular lattice contacts. CONCLUSIONS: The first nine helices adopt a fold similar to that observed for RGS proteins, although the sequence identity with other such known structures is below 20%. The last three helices are an integral extension of the RGS fold, packing tightly against helices 3 and 4 with multiple hydrophobic interactions. Comparison with RGS proteins suggests features that are likely relevant for interaction with G proteins. Finally, we conclude that the strategy used to produce crystals was beneficial and might be applicable to other proteins resistant to crystallization.

Selected figure(s)

Figure 4.
Figure 4. Tertiary Fold of the RGSL Domain(a) This ribbon diagram is colored from the N terminus (red) to the C terminus (blue), and secondary structural elements are numbered sequentially.(b) Alignment of the RGSL domain and the structure of RGS4 (gray) is shown in stereo as an a-carbon trace from the same perspective as in (a). The RGSL is colored green for the overlapping portion and colored magenta for the C-terminal extension. The residue numbering of the RGSL domain is labeled according to the full-length PDZ-RhoGEF

The above figure is reprinted by permission from Cell Press: Structure (2001, 9, 559-569) copyright 2001.

Figure was selected by an automated process.