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Dynamic properties of the Ras switch I region and its importance for binding to effectors.

Spoerner M, Herrmann C, Vetter IR, Kalbitzer HR, Wittinghofer A
Proc Natl Acad Sci U S A 98 4944-9 (2001)
Cited: 174 times
EuropePMC logo PMID: 11320243

Abstract

We have investigated the dynamic properties of the switch I region of the GTP-binding protein Ras by using mutants of Thr-35, an invariant residue necessary for the switch function. Here we show that these mutants, previously used as partial loss-of-function mutations in cell-based assays, have a reduced affinity to Ras effector proteins without Thr-35 being involved in any interaction. The structure of Ras(T35S)(.)GppNHp was determined by x-ray crystallography. Whereas the overall structure is very similar to wildtype, residues from switch I are completely invisible, indicating that the effector loop region is highly mobile. (31)P-NMR data had indicated an equilibrium between two rapidly interconverting conformations, one of which (state 2) corresponds to the structure found in the complex with the effectors. (31)P-NMR spectra of Ras mutants (T35S) and (T35A) in the GppNHp form show that the equilibrium is shifted such that they occur predominantly in the nonbinding conformation (state 1). On addition of Ras effectors, Ras(T35S) but not Ras(T35A) shift to positions corresponding to the binding conformation. The structural data were correlated with kinetic experiments that show two-step binding reaction of wild-type and (T35S)Ras with effectors requires the existence of a rate-limiting isomerization step, which is not observed with T35A. The results indicate that minor changes in the switch region, such as removing the side chain methyl group of Thr-35, drastically affect dynamic behavior and, in turn, interaction with effectors. The dynamics of the switch I region appear to be responsible for the conservation of this threonine residue in GTP-binding proteins.

Articles - 1iaq mentioned but not cited (4)

  1. Dynamic properties of the Ras switch I region and its importance for binding to effectors. Spoerner M, Herrmann C, Vetter IR, Kalbitzer HR, Wittinghofer A. Proc Natl Acad Sci U S A 98 4944-4949 (2001)
  2. Distance matrix-based approach to protein structure prediction. Kloczkowski A, Jernigan RL, Wu Z, Song G, Yang L, Kolinski A, Pokarowski P. J Struct Funct Genomics 10 67-81 (2009)
  3. A structural model of a Ras-Raf signalosome. Mysore VP, Zhou ZW, Ambrogio C, Li L, Kapp JN, Lu C, Wang Q, Tucker MR, Okoro JJ, Nagy-Davidescu G, Bai X, Plückthun A, Jänne PA, Westover KD, Shan Y, Shaw DE. Nat Struct Mol Biol 28 847-857 (2021)
  4. Organization of Farnesylated, Carboxymethylated KRAS4B on Membranes. Barklis E, Stephen AG, Staubus AO, Barklis RL, Alfadhli A. J Mol Biol 431 3706-3717 (2019)


Reviews citing this publication (25)

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  2. Direct small-molecule inhibitors of KRAS: from structural insights to mechanism-based design. Ostrem JM, Shokat KM. Nat Rev Drug Discov 15 771-785 (2016)
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  15. The Ins and Outs of RAS Effector Complexes. Kiel C, Matallanas D, Kolch W. Biomolecules 11 236 (2021)
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  18. Current status of the development of Ras inhibitors. Shima F, Matsumoto S, Yoshikawa Y, Kawamura T, Isa M, Kataoka T. J Biochem 158 91-99 (2015)
  19. Dynamically encoded reactivity of Ras enzymes: opening new frontiers for drug discovery. Pálfy G, Menyhárd DK, Perczel A. Cancer Metastasis Rev 39 1075-1089 (2020)
  20. Exploiting RAS Nucleotide Cycling as a Strategy for Drugging RAS-Driven Cancers. Mattox TE, Chen X, Maxuitenko YY, Keeton AB, Piazza GA. Int J Mol Sci 21 E141 (2019)
  21. KRAS-Mutant Non-Small-Cell Lung Cancer: From Past Efforts to Future Challenges. Ceddia S, Landi L, Cappuzzo F. Int J Mol Sci 23 9391 (2022)
  22. CRIB effector disorder: exquisite function from chaos. Owen D, Mott HR. Biochem Soc Trans 46 1289-1302 (2018)
  23. Relevance of the kinetic equilibrium of forces to the control of the cell cycle by Ras proteins. Becker EW. Biol Chem 385 41-47 (2004)
  24. Peptides That Block RAS-p21 Protein-Induced Cell Transformation. Pincus MR, Lin B, Patel P, Gabutan E, Zohar N, Bowne WB. Biomedicines 11 471 (2023)
  25. Targeting small GTPases: emerging grasps on previously untamable targets, pioneered by KRAS. Yin G, Huang J, Petela J, Jiang H, Zhang Y, Gong S, Wu J, Liu B, Shi J, Gao Y. Signal Transduct Target Ther 8 212 (2023)

Articles citing this publication (145)



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