1RRB

THE RAS-BINDING DOMAIN OF RAF-1 FROM RAT, NMR, 1 STRUCTURE

Summary for 1RRB

• Entry DOI: 10.2210/pdb1rrb/pdb
• Descriptor: RAF PROTO-ONCOGENE SERINE/THREONINE-PROTEIN KINASE (1 entity in total)
• Functional Keywords: raf-1, ras-binding domain, transferase, serine/threonine-protein kinase, riken structural genomics/proteomics initiative, rsgi, structural genomics
• Biological source: Rattus norvegicus (Norway rat)
• Cellular location: Cytoplasm: P11345
• Total number of polymer chains: 1
• Total formula weight: 11960.82

Authors

Terada, T.,Ito, Y.,Shirouzu, M.,Tateno, M.,Hashimoto, K.,Kigawa, T.,Ebisuzaki, T.,Takio, K.,Shibata, T.,Yokoyama, S.,Smith, B.O.,Laue, E.D.,Cooper, J.A.,RIKEN Structural Genomics/Proteomics Initiative (RSGI) (deposition date: 1998-03-26, release date: 1999-03-30, Last modification date: 2024-05-22)

Primary citation

Terada, T.,Ito, Y.,Shirouzu, M.,Tateno, M.,Hashimoto, K.,Kigawa, T.,Ebisuzaki, T.,Takio, K.,Shibata, T.,Yokoyama, S.,Smith, B.O.,Laue, E.D.,Cooper, J.A.
Nuclear magnetic resonance and molecular dynamics studies on the interactions of the Ras-binding domain of Raf-1 with wild-type and mutant Ras proteins.
J.Mol.Biol., 286:219-232, 1999

PubMed Abstract: The Ras protein and its homolog, Rap1A, have an identical "effector region" (residues 32-40) preceded by Asp30-Glu31 and Glu30-Lys31, respectively. In the complex of the "Ras-like" E30D/K31E mutant Rap1A with the Ras-binding domain (RBD), residues 51-131 of Raf-1, Glu31 in Rap1A forms a tight salt bridge with Lys84 in Raf-1. However, we have recently found that Raf-1 RBD binding of Ras is indeed reduced by the E31K mutation, but is not affected by the E31A mutation. Here, the "Rap1A-like" D30E/E31K mutant of Ras was prepared and shown to bind the Raf-1 RBD less strongly than wild-type Ras, but slightly more tightly than the E31K mutant. The backbone 1H, 13C, and 15N magnetic resonances of the Raf-1 RBD were assigned in complexes with the wild-type and D30E/E31K mutant Ras proteins in the guanosine 5'-O-(beta,gamma-imidotriphosphate)-bound form. The Lys84 residue in the Raf-1 RBD exhibited a large change in chemical shift upon binding wild-type Ras, suggesting that Lys84 interacts with wild-type Ras. The D30E/E31K mutant of Ras caused nearly the same perturbations in Raf-1 chemical shifts, including that of Lys84. We hypothesized that Glu31 in Ras may not be the major salt bridge partner of Lys84 in Raf-1. A molecular dynamics simulation of a model structure of the Raf-1 RBD.Ras.GTP complex suggested that Lys84 in Raf-1 might instead form a tight salt bridge with Asp33 in Ras. Consistent with this, the D33A mutation in Ras greatly reduced its Raf-I RBD binding activity. We conclude that the major salt bridge partner of Lys84 in Raf-1 may be Asp33 in Ras.

PubMed: 9931261
DOI: 10.1006/jmbi.1998.2472

Experimental method

SOLUTION NMR