PDBsum entry 2erz

Transferase

PDB id: 2erz

Contents

Protein chains

336 a.a. *

20 a.a. *

Ligands

HFS

Waters ×61

* Residue conservation analysis

PDB id:

2erz

Name:

Transferase

Title:

Crystal structure of c-amp dependent kinase (pka) bound to hydroxyfasudil

Structure:

Camp-dependent protein kinase, alpha-catalytic subunit. Chain: e. Synonym: pka c-alpha. Engineered: yes. Camp-dependent protein kinase inhibitor, alpha form. Chain: i. Fragment: residues 5-24. Synonym: pki-alpha, camp- dependent protein kinase inhibitor, muscle/brain isoform.

Source:

Mus musculus. House mouse. Organism_taxid: 10090. Gene: prkaca, pkaca. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Other_details: this sequence occurs naturally in oryctolagus cuniculus (rabbit)

Biol. unit:

Dimer (from PQS)

Resolution:

2.20Å R-factor: 0.227 R-free: 0.287

Authors:

M.Jacobs

Key ref:

M.Jacobs et al. (2006). The structure of dimeric ROCK I reveals the mechanism for ligand selectivity. J Biol Chem, 281, 260-268. PubMed id: 16249185 DOI: 10.1074/jbc.M508847200

Date:

25-Oct-05 Release date: 08-Nov-05

Protein chain

P05132  (KAPCA_MOUSE) -  cAMP-dependent protein kinase catalytic subunit alpha from Mus musculus

Seq: 351 a.a.

Struc: 336 a.a.*

Protein chain

P61926  (IPKA_RABIT) -  cAMP-dependent protein kinase inhibitor alpha from Oryctolagus cuniculus

Seq: 76 a.a.

Struc: 20 a.a.

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

Enzyme reactions

• Enzyme class: Chain E: E.C.2.7.11.11 - cAMP-dependent protein kinase.
• Reaction:
  1. L-seryl-[protein] + ATP = O-phospho-L-seryl-[protein] + ADP + H⁺
  2. L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] + ADP + H⁺

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

reference

DOI no: 10.1074/jbc.M508847200

J Biol Chem 281:260-268 (2006)

PubMed id: 16249185

The structure of dimeric ROCK I reveals the mechanism for ligand selectivity.

M.Jacobs, K.Hayakawa, L.Swenson, S.Bellon, M.Fleming, P.Taslimi, J.Doran.

ABSTRACT

ROCK or Rho-associated kinase, a serine/threonine kinase, is an effector of Rho-dependent signaling and is involved in actin-cytoskeleton assembly and cell motility and contraction. The ROCK protein consists of several domains: an N-terminal region, a kinase catalytic domain, a coiled-coil domain containing a RhoA binding site, and a pleckstrin homology domain. The C-terminal region of ROCK binds to and inhibits the kinase catalytic domains, and this inhibition is reversed by binding RhoA, a small GTPase. Here we present the structure of the N-terminal region and the kinase domain. In our structure, two N-terminal regions interact to form a dimerization domain linking two kinase domains together. This spatial arrangement presents the kinase active sites and regulatory sequences on a common face affording the possibility of both kinases simultaneously interacting with a dimeric inhibitory domain or with a dimeric substrate. The kinase domain adopts a catalytically competent conformation; however, no phosphorylation of active site residues is observed in the structure. We also determined the structures of ROCK bound to four different ATP-competitive small molecule inhibitors (Y-27632, fasudil, hydroxyfasudil, and H-1152P). Each of these compounds binds with reduced affinity to cAMP-dependent kinase (PKA), a highly homologous kinase. Subtle differences exist between the ROCK- and PKA-bound conformations of the inhibitors that suggest that interactions with a single amino acid of the active site (Ala215 in ROCK and Thr183 in PKA) determine the relative selectivity of these compounds. Hydroxyfasudil, a metabolite of fasudil, may be selective for ROCK over PKA through a reversed binding orientation.

Selected figure(s)

Figure 1.
FIGURE 1. Overall structure of the ROCK/Y-27632 complex. A, the ROCK protein dimer is drawn with -sheets as arrows and -helices as cylinders. One monomer is drawn in gray, and the others are colored by protein region. The N-terminal dimerization domain is shown in red. The N-terminal kinase domain (dark blue) is shown with the glycine-rich loop drawn in green. The hinge connecting the two domains is colored orange. The C-terminal kinase domain is shown in light blue with the activation loop in purple and the kinase tail in yellow. Y-27632 is shown in the active site near the glycine-rich loop and the hinge. B, a surface representation of the dimer is shown where both monomers are colored by region. Y-27632 is shown in the active site as spheres. A model of the substrate peptide is shown as a pink cylinder and strand, based upon a superposition of the ROCK structure and the PKA/ATP/peptide complex (Protein Data Bank code 1ATP [PDB] ). C and D, an expanded view of the dimerization domain is shown in two orientations, differing by a 90° rotation. All of the structure figures were made with PYMOL (58).

Figure 3.
FIGURE 3. Phosphothreonine-binding site in PKA aligned with ROCK. The activation loops of PKA (gray) and ROCK (green) are shown with interactions between side chains and the phosphate shown as purple dotted lines.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2006, 281, 260-268) copyright 2006.

Figures were selected by an automated process.