Cited by 149Protein Kinase Regulation and GTPase SignalingProtein Structure and DynamicsReceptor Mechanisms and Signaling

Conformational states of human rat sarcoma (RAS) protein complexed with its natural ligand GTP and their role for effector interaction and GTP hydrolysis

Open access
Spoerner, Michael, Hozsa, Constantin, Poetzl, Johann A., Reiss, Kerstin, Ganser, Petra, Geyer, Matthias, Kalbitzer, Hans Robert·2010·Journal of Biological Chemistry

Abstract

The guanine nucleotide-binding protein Ras exists in solution in two different conformational states when complexed with different GTP analogs such as GppNHp or GppCH(2)p. State 1 has only a very low affinity to effectors and seems to be recognized by guanine nucleotide exchange factors, whereas state 2 represents the high affinity effector binding state. In this work we investigate Ras in complex with the physiological nucleoside triphosphate GTP. By polarization transfer (31)P NMR experiments and effector binding studies we show that Ras(wt)·Mg(2+)·GTP also exists in a dynamical equilibrium between the weakly populated conformational state 1 and the dominant state 2. At 278 K the equilibrium constant between state 1 and state 2 of C-terminal truncated wild-type Ras(1-166) K(12) is 11.3. K(12) of full-length Ras is >20, suggesting that the C terminus may also have a regulatory effect on the conformational equilibrium. The exchange rate (k(ex)) for Ras(wt)·Mg(2+)·GTP is 7 s(-1) and thus 18-fold lower compared with that found for the Ras·GppNHp complex. The intrinsic GTPase activity substantially increases after effector binding for the switch I mutants Ras(Y32F), (Y32R), (Y32W), (Y32C/C118S), (T35S), and the switch II mutant Ras(G60A) by stabilizing state 2, with the largest effect on Ras(Y32R) with a 13-fold increase compared with wild-type. In contrast, no acceleration was observed in Ras(T35A). Thus Ras in conformational state 2 has a higher affinity to effectors as well as a higher GTPase activity. These observations can be used to explain why many mutants have a low GTPase activity but are not oncogenic.

Integrity signals

Registry

Crossref · 10.1074/jbc.m110.145235

Cites

43 of 43 references verified0 citation errors · 0 not checkable

Statistics

Recomputing p-values needs the paper’s full text.

Claims the paper makes

We couldn’t retrieve this paper’s full text, so its claims and self-check weren’t run. The registry and citation checks still apply.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Cites

Flagged first, then by citations

Cited by

Sorted by citations

Authors
Spoerner, M., Hozsa, C., Poetzl, J. A., Reiss, K., Ganser, P., Geyer, M., & Kalbitzer, H. R.
Affiliations
  • University of Regensburg (DE)
  • Max Planck Society (DE)
Corresponding author
Kalbitzer, Hans Robert
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2010-12
Volume
285
Issue
51
Pages
39768-39778
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2024944232
Cited by
148
References
43
Citations by year
2026: 3 · 2025: 11 · 2024: 12 · 2023: 9 · 2022: 9 · 2021: 6 · 2020: 14 · 2019: 9 · 2018: 10 · 2017: 11 · 2016: 6 · 2015: 12 · 2014: 8 · 2013: 10 · 2012: 11
FWCI
3.65
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Max Planck Institute for Plasma Physics (submittedVersion)
  • University of Regensburg Publication Server (University of Regensburg) (submittedVersion)
  • MPG.PuRe (Max Planck Society) (submittedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
Keywords
Ras proteins, GTP binding, conformational equilibrium, effector binding, GTPase activity, guanine nucleotide exchange factor, oncogenicity, Gpp(NH)p, 31P NMR
MeSH terms
Animals, Guanosine Triphosphate, Humans, Proto-Oncogene Proteins p21(ras), Protein Structure, Tertiary, Nuclear Magnetic Resonance, Biomolecular, Amino Acid Substitution, Mutation, Missense, Rats

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review