PDBsum entry 2fnf

Apoptosis

PDB id: 2fnf

Contents

Protein chain

59 a.a. *

Metals

_ZN ×2

* Residue conservation analysis

PDB id:

2fnf

Name:

Apoptosis

Title:

C1 domain of nore1

Structure:

Putative ras effector nore1. Chain: x. Fragment: cysteine rich domain. Engineered: yes

Source:

Mus musculus. House mouse. Organism_taxid: 10090. Expressed in: escherichia coli. Expression_system_taxid: 562.

NMR struc:

30 models

Authors:

E.Harjes,S.Harjes,S.Wohlgemuth,E.Krieger,C.Herrmann,K.H.Muller, P.Bayer

Key ref:

E.Harjes et al. (2006). GTP-Ras disrupts the intramolecular complex of C1 and RA domains of Nore1. Structure, 14, 881-888. PubMed id: 16698549 DOI: 10.1016/j.str.2006.03.008

Date:

11-Jan-06 Release date: 07-Feb-06

Protein chain

Q5EBH1  (RASF5_MOUSE) -  Ras association domain-containing protein 5 from Mus musculus

Seq: 413 a.a.

Struc: 59 a.a.

Enzyme reactions

• Enzyme class: E.C.?

DOI no: 10.1016/j.str.2006.03.008

Structure 14:881-888 (2006)

PubMed id: 16698549

GTP-Ras disrupts the intramolecular complex of C1 and RA domains of Nore1.

E.Harjes, S.Harjes, S.Wohlgemuth, K.H.Müller, E.Krieger, C.Herrmann, P.Bayer.

ABSTRACT

The novel Ras effector mNore1, capable of inducing apoptosis, is a multidomain protein. It comprises a C1 domain homologous to PKC and an RA domain similar to the Ras effectors AF-6 and RalGDS. Here, we determine the affinity of these two domains to the active forms of Ras and Rap1 using isothermal calorimetric titration. The interaction of Ras/Rap1-GTP with the RA domain of mNore1 is weakened significantly by direct binding of the C1 domain to the RA domain. In order to analyze this observation in atomic detail, we solved the C1 solution structure by NMR. By determining chemical shifts and relaxation rates, we can show an intramolecular complex of C1-RA. GTP-Ras titration and binding to RA disrupts this complex and displaces the C1 domain. Once the C1 domain tumbles freely in solution, a lipid binding interface becomes accessible. Furthermore, we provide evidence of phosphatidylinositol 3-phosphate binding of the free C1 domain.

Selected figure(s)

Figure 6.
Figure 6. Comparison of Relaxation Data
Relaxation rates were measured for mNore1-C1 (squares), the RA-C1 construct (triangles), and the Ras-GppNHp-titrated RA-C1 construct (circles). The R[2]/R[1] quotient was plotted versus the corresponding amino acid. Figure 6. Comparison of Relaxation DataRelaxation rates were measured for mNore1-C1 (squares), the RA-C1 construct (triangles), and the Ras-GppNHp-titrated RA-C1 construct (circles). The R[2]/R[1] quotient was plotted versus the corresponding amino acid.

Figure 8.
Figure 8. Lipid Binding Specificity of the mNore1-C1 Domain
A total of 26 different lipids on a nitrocellulose membrane were probed with recombinant GST-mNore1-C1 (GST-95-166), were subsequently labeled with anti-GST-antibody and HRP-coupled anti-mouse antibody, and were followed by ECL detection. Figure 8. Lipid Binding Specificity of the mNore1-C1 DomainA total of 26 different lipids on a nitrocellulose membrane were probed with recombinant GST-mNore1-C1 (GST-95-166), were subsequently labeled with anti-GST-antibody and HRP-coupled anti-mouse antibody, and were followed by ECL detection.

The above figures are reprinted by permission from Cell Press: Structure (2006, 14, 881-888) copyright 2006.

Figures were selected by an automated process.