PDBsum entry 1s1c

Signaling protein

PDB id: 1s1c

Contents

Protein chains

179 a.a. *

69 a.a. *

Ligands

GNP ×2

Metals

_MG ×2

Waters ×55

* Residue conservation analysis

PDB id:

1s1c

Name:

Signaling protein

Title:

Crystal structure of the complex between the human rhoa and rho- binding domain of human rocki

Structure:

Transforming protein rhoa. Chain: a, b. Fragment: rhoa. Synonym: h12. Engineered: yes. Rho-associated, coiled-coil containing protein kinase 1. Chain: x, y. Fragment: rho-binding domain of rocki, residues 947-1015. Synonym: p160rock, p160-rock.

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: arha, arh12, rhoa, rho12. Expressed in: escherichia coli. Expression_system_taxid: 562. Expression_system_taxid: 562

Biol. unit:

Tetramer (from PQS)

Resolution:

2.60Å R-factor: 0.219 R-free: 0.257

Authors:

R.Dvorsky,L.Blumenstein,I.R.Vetter,M.R.Ahmadian

Key ref:

R.Dvorsky et al. (2004). Structural insights into the interaction of ROCKI with the switch regions of RhoA. J Biol Chem, 279, 7098-7104. PubMed id: 14660612 DOI: 10.1074/jbc.M311911200

Date:

06-Jan-04 Release date: 10-Feb-04

Protein chains

P61586  (RHOA_HUMAN) -  Transforming protein RhoA from Homo sapiens

Seq: 193 a.a.

Struc: 179 a.a.

Protein chains

Q13464  (ROCK1_HUMAN) -  Rho-associated protein kinase 1 from Homo sapiens

Seq: 1354 a.a.

Struc: 69 a.a.*

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

Enzyme reactions

• Enzyme class 1: Chains A, B: E.C.3.6.5.2 - small monomeric GTPase.
• Reaction: GTP + H2O = GDP + phosphate + H⁺

GTP + H2O = GDP (Bound ligand (Het Group name = GNP) matches with 81.82% similarity) + phosphate + H(+)

• Enzyme class 2: Chains X, Y: E.C.2.7.11.1 - non-specific serine/threonine 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⁺

L-seryl-[protein] + ATP = O-phospho-L-seryl-[protein] (Bound ligand (Het Group name = GNP) matches with 78.79% similarity) + ADP + H(+)

L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] (Bound ligand (Het Group name = GNP) matches with 78.79% similarity) + ADP + H(+)

Note, where more than one E.C. class is given (as above), each may correspond to a different protein domain or, in the case of polyprotein precursors, to a different mature protein.

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

reference

DOI no: 10.1074/jbc.M311911200

J Biol Chem 279:7098-7104 (2004)

PubMed id: 14660612

Structural insights into the interaction of ROCKI with the switch regions of RhoA.

R.Dvorsky, L.Blumenstein, I.R.Vetter, M.R.Ahmadian.

ABSTRACT

The Rho-ROCK pathway modulates the phosphorylation level of a variety of important signaling proteins and is thereby involved in miscellaneous cellular processes including cell migration, neurite outgrowth, and smooth muscle contraction. The observation of the involvement of the Rho-ROCK pathway in tumor invasion and in diseases such as hypertension and bronchial asthma makes it an interesting target for drug development. We herein present the crystal structure of the complex between active RhoA and the Rho-binding domain of ROCKI. The Rho-binding domain structure forms a parallel alpha-helical coiled-coil dimer and, in contrast to the published Rho-protein kinase N structure, binds exclusively to the switch I and II regions of the guanosine 5'-(beta,gamma-imido)triphosphate-bound RhoA. The switch regions of two different RhoA molecules form a predominantly hydrophobic patch, which is complementarily bound by two identical short helices of 13 residues (amino acids 998-1010). The identified ROCK-binding site of RhoA strikingly supports the assumption of a common consensus-binding site for effector recognition.

Selected figure(s)

Figure 3.
FIG. 3. Determinants of the ROCK specificity toward Rho. A, stereoview of superimposed structures of RhoA (orange) (33), Rac1 (yellow) (50), and Cdc42 (brown) (51), focusing on the ROCK-binding region of RhoA. Amino acids are labeled according to the standard numbering of RhoA. B, surface representation of the GTPases in the same orientation as in A. The residues that interact with ROCKI-RBD are colored as follows: blue, positively charged nitrogen atoms; red, negatively charged oxygen atoms; green, non-charged hydrophobic atoms; black, the oxygen atom of Tyr-66. Phe-39, found to be crucial for the specificity toward ROCKI, is shown in yellow.

Figure 4.
FIG. 4. ROCK and PKN contact sites of RhoA. The contact site I proposed as the main contact site of RhoA with PKN (27) is colored in brown. The common binding site of RhoA for both ROCKI and PKN (contact site II) is highlighted in orange. The sites interacting exclusively with PKN and ROCK are colored in yellow and red, respectively. All of the contact sites were determined using the 4.5-Å threshold.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2004, 279, 7098-7104) copyright 2004.

Figures were selected by an automated process.