2ETR

Crystal Structure of ROCK I bound to Y-27632

Summary for 2ETR

• Entry DOI: 10.2210/pdb2etr/pdb
• Related: 2ERZ 2ESM 2ETK 2ETO
• Descriptor: Rho-associated protein kinase 1, (R)-TRANS-4-(1-AMINOETHYL)-N-(4-PYRIDYL) CYCLOHEXANECARBOXAMIDE (3 entities in total)
• Functional Keywords: dimerization, dimer, phosphorylation, kinase, yoshitomi, transferase
• Biological source: Homo sapiens (human)
• Cellular location: Cytoplasm: Q13464
• Total number of polymer chains: 2
• Total formula weight: 96519.24

Authors

Jacobs, M. (deposition date: 2005-10-27, release date: 2005-11-08, Last modification date: 2023-08-23)

Primary citation

Jacobs, M.,Hayakawa, K.,Swenson, L.,Bellon, S.,Fleming, M.,Taslimi, P.,Doran, J.
The Structure of Dimeric ROCK I Reveals the Mechanism for Ligand Selectivity.
J.Biol.Chem., 281:260-268, 2006

PubMed 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.

PubMed: 16249185
DOI: 10.1074/jbc.M508847200

Experimental method

X-RAY DIFFRACTION (2.6 Å)