PDBsum entry: 1omw

Transferase

PDB id: 1omw

Contents

Protein chains

614 a.a. *

339 a.a. *

61 a.a. *

Waters ×26

* Residue conservation analysis

PDB id:

1omw

Name:

Transferase

Title:

Crystal structure of the complex between g protein-coupled r kinase 2 and heterotrimeric g protein beta 1 and gamma 2 su

Structure:

G-protein coupled receptor kinase 2. Chain: a. Synonym: grk2, beta-ark-1, beta-adrenergic receptor kinase engineered: yes. Mutation: yes. Guanine nucleotide-binding protein g(i)/g(s)/g(t) subunit 1. Chain: b. Synonym: transducin beta chain 1.

Source:

Bos taurus. Cattle. Organism_taxid: 9913. Gene: grk2. Expressed in: spodoptera frugiperda. Expression_system_taxid: 7108. Gene: gnb1. Gene: gng2.

Biol. unit:

Trimer (from PQS)

Resolution:

2.50Å R-factor: 0.216 R-free: 0.252

Authors:

D.T.Lodowski,J.A.Pitcher,W.D.Capel,R.J.Lefkowitz,J.J.G.Tesme

Key ref:

D.T.Lodowski et al. (2003). Keeping G proteins at bay: a complex between G protein-coupled receptor kinase 2 and Gbetagamma. Science, 300, 1256-1262. PubMed id: 12764189 DOI: 10.1126/science.1082348

Date:

26-Feb-03 Release date: 03-Jun-03

Protein chain

P21146  (ARBK1_BOVIN) -  Beta-adrenergic receptor kinase 1

Seq: 689 a.a.

Struc: 614 a.a.

Protein chain

P62871  (GBB1_BOVIN) -  Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1

Seq: 340 a.a.

Struc: 339 a.a.

Protein chain

P63212  (GBG2_BOVIN) -  Guanine nucleotide-binding protein G(I)/G(S)/G(O) subunit gamma-2

Seq: 71 a.a.

Struc: 61 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: Chain A: E.C.2.7.11.15 - [Beta-adrenergic-receptor] kinase.
• Reaction: ATP + [beta-adrenergic receptor] = ADP + [beta-adrenergic receptor] phosphate

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

 Gene Ontology (GO) functional annotation 

• Cellular component: intracellular (8 terms)
• Biological process: termination of G-protein coupled receptor signaling pathway (23 terms)
• Biochemical function: nucleotide binding (15 terms)

reference

DOI no: 10.1126/science.1082348

Science 300:1256-1262 (2003)

PubMed id: 12764189

Keeping G proteins at bay: a complex between G protein-coupled receptor kinase 2 and Gbetagamma.

D.T.Lodowski, J.A.Pitcher, W.D.Capel, R.J.Lefkowitz, J.J.Tesmer.

ABSTRACT

The phosphorylation of heptahelical receptors by heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptor kinases (GRKs) is a universal regulatory mechanism that leads to desensitization of G protein signaling and to the activation of alternative signaling pathways.We determined the crystallographic structure of bovine GRK2 in complex with G protein beta1gamma2 subunits.Our results show how the three domains of GRK2-the RGS (regulator of G protein signaling) homology, protein kinase, and pleckstrin homology domains-integrate their respective activities and recruit the enzyme to the cell membrane in an orientation that not only facilitates receptor phosphorylation, but also allows for the simultaneous inhibition of signaling by Galpha and Gbetagamma subunits.

Selected figure(s)

Figure 1.
Fig. 1. Quaternary structure of the GRK2-Gß[1] [2] complex. (A) Membrane-proximal view of the GRK2-Gß complex. The RH (RGS homology) domain of GRK2 is colored violet. The kinase domain is depicted with yellow helices and olive-green ß strands. The PH domain is tan, Gß[1] blue, and G [2] green. The long axis of the RH domain is declined from the center of the enzyme into the page by about 45°. The first and last observed residues of GRK2 are labeled "N" (residue 29) and "C" (residue 668), respectively. The C-terminal residue of G (Cys68) is labeled "geranyl" to indicate the site of geranylgeranylation. Connections of disordered loops in GRK2 are annotated as follows: I and I' correspond to residues 475 and 496, respectively, of the kinase domain; II and II' to residues 569 and 576, respectively, of the PH domain. (B) Side view of the GRK2-Gß complex, rotated 90° around a horizontal axis from the view in (A). The flat, membrane-proximal surface spans the top of the complex. (C) Electrostatic surface potential of the membrane-proximal surface of the complex (21). The orientation is the same as in (A). Basic regions are colored blue, acidic regions red, and neutral regions white. (D) Electrostatic surface potential of the GRK2-Gß complex in the same orientation as (B).

Figure 5.
Fig. 5. The GRK2 PH domain and its interface with Gß . (A) The GRK2 PH domain. Residues implicated in the binding of anionic phospholipids (14, 46) are drawn with blue side chains. For perspective, inositol 1,4,5-trisphosphate [Ins(1,4,5)P[3]] is modeled from the structure of PLC (60). Two residues from GRK2, Lys567 and Arg579, are in position to coordinate the phosphates of the anionic head group, as do equivalent residues in other PH domains (45). The four regions within the primary sequence of the GRK2 PH domain that contact Gß are each drawn witha different color for reference [see (B) and (D)] (21). These regions form a continuous surface that includes the ß1-ß4 sheet of the PH domain, the extended portion of the CT helix, and the C-terminal tail. The conformation of the ß1-ß4 sheet is highly conserved among PH domains of known structure (table S1). Therefore, many PH domains have a surface that is complementary in shape to the effector-binding surface of Gß . (B) Specific interactions between the PH domain and Gß. The location of each interacting residue within the tertiary structure of the PH domain is indicated alongside each amino acid. In addition to the interactions shown, Arg689 of GRK2, which was not modeled because of weak electron density, is also expected to contact the surface of Gß . (C) Comparison of the surfaces of Gß that bind G subunits and GRK2. The molecular surface of Gß was colored according to its contacts with G (blue), GRK2 (red), or neither (white). Common binding surfaces are colored purple. The footprint of G on the surface of Gß overlaps extensively with that of the GRK2 PH domain, and thus their binding is mutually exclusive. Positions of various residues from Gß that contact the PH domain of GRK2 are labeled for reference. (D) Stereoview of the PH domain-Gß[1] interface. Residues from Gß are drawn with gray carbons, residues from the PH domain with tan carbons. Hydrogen bonds or salt bridges between residues are indicated with dashed lines. The side chain of Met664 from the RH domain binds within a hydrophobic pocket, one wall of which is formed by Leu117 of Gß (omitted for clarity). Trp99 of Gß docks into a hydrophobic groove at the interface such that its indole nitrogen is oriented toward solvent. Although electron density for the side chain of Lys663 is not observed beyond Cß, it could extend far enough into the central channel of Gß to allow interaction of its N atom witha ring of seven carbonyl oxygens donated by the innermost strand from each blade of Gß. Figure S4 details additional interactions between the PH domain C-terminal tail and Gß.

The above figures are reprinted by permission from the AAAs: Science (2003, 300, 1256-1262) copyright 2003.

Figures were selected by the author.