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PDBsum entry 2fnf
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References listed in PDB file
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Key reference
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Title
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Gtp-Ras disrupts the intramolecular complex of c1 and ra domains of nore1.
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Authors
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E.Harjes,
S.Harjes,
S.Wohlgemuth,
K.H.Müller,
E.Krieger,
C.Herrmann,
P.Bayer.
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Ref.
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Structure, 2006,
14,
881-888.
[DOI no: ]
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PubMed id
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Note: In the PDB file this reference is
annotated as "TO BE PUBLISHED". The citation details given above have
been manually determined.
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Abstract
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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.
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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.
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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.
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The above figures are
reprinted
by permission from Cell Press:
Structure
(2006,
14,
881-888)
copyright 2006.
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