The DOI matches its Crossref record; 83 of the 84 references it cites were found in a registry or an index.
The small GTPases K-Ras, N-Ras, and H-Ras have distinct biochemical properties determined by allosteric effects
Open accessAbstract
H-Ras, K-Ras, and N-Ras are small GTPases that are important in the control of cell proliferation, differentiation, and survival, and their mutants occur frequently in human cancers. The G-domain, which catalyzes GTP hydrolysis and mediates downstream signaling, is 95% conserved between the Ras isoforms. Because of their very high sequence identity, biochemical studies done on H-Ras have been considered representative of all three Ras proteins. We show here that this is not a valid assumption. Using enzyme kinetic assays under identical conditions, we observed clear differences between the three isoforms in intrinsic catalysis of GTP by Ras in the absence and presence of the Ras-binding domain (RBD) of the c-Raf kinase protein (Raf-RBD). Given their identical active sites, isoform G-domain differences must be allosteric in origin, due to remote isoform-specific residues that affect conformational states. We present the crystal structure of N-Ras bound to a GTP analogue and interpret the kinetic data in terms of structural features specific for H-, K-, and N-Ras.
Overview
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
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?
In the citing paper
“RAS mutants were once viewed as being ‘locked’ in the GTP-bound ‘on’ state, impervious to GAP-stimulated or intrinsic GTP hydrolysis, and hence undruggable. However, detailed structural and biochemical analyses of various RAS mutants associated with cancer and ‘RASopathies’”
From The current state of the art and future trends in RAS-targeted cancer therapies (2022) · cites this paper as [45]
In the citing paper
“At the cellular level, the isoform specificity to effector proteins is primarily determined via membrane interactions [24], but the differences among RAS isoforms’ absolute effector protein binding affinities rise from allosteric effects [25].”
Citations
Cites
Bradford, M. M. (1976) · Analytical Biochemistry · doi:10.1016/0003-2697(76)90527-3 · Cited by 255,302 · cited 1×
“Protein concentrations for biochemical experiments were determined using the Bradford assay just prior to use (79).”
MethodBerman, H. M. (2000) · Nucleic Acids Research · doi:10.1093/nar/28.1.235 · Cited by 40,196
[20] Processing of X-ray diffraction data collected in oscillation mode
Otwinowski, Z., & Minor, W. (1997) · Methods in Enzymology · doi:10.1016/s0076-6879(97)76066-x · Cited by 33,475
PHENIX : a comprehensive Python-based system for macromolecular structure solution
Adams, P. D., Afonine, P. V., Bunkóczi, G., Chen, V. B., Davis, I. W., Echols, N., Headd, J. J., Hung, L.-W., Kapral, G. J., Grosse-Kunstleve, R. W., et al. (2010) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444909052925 · Cited by 23,716 · cited 1×
“Molecular replacement and structure refinement were performed using the PHENIX suite of programs (81) with the H-Ras-GppNHp coordinates with PDB code 1CTQ used as the initial phasing model.”
MethodNMRPipe: A multidimensional spectral processing system based on UNIX pipes
Delaglio, F., Grzesiek, S., Vuister, G. W., Zhu, G., Pfeifer, J., & Bax, A. (1995) · Journal of Biomolecular NMR · doi:10.1007/bf00197809 · Cited by 15,961
PHENIX: a comprehensive Python-based system for macromolecular structure solution
Adams, P. D., Afonine, P. V., Bunkòczi, G., Chen, V. B., Davis, I., Echols, N., Headd, J. J., Hung, L., Kapral, G. J., Grosse‐Kunstleve, R. W., et al. (2012) · International Tables for Crystallography · doi:10.1107/97809553602060000865 · Cited by 7,334
Nonlinear Regression Analysis and Its Applications
Bates, D. M., & Watts, D. G. (1988) · Wiley series in probability and statistics · doi:10.1002/9780470316757 · Cited by 4,017
The GTPase superfamily: conserved structure and molecular mechanism
Bourne, H. R., Sanders, D. A., & McCormick, F. (1991) · Nature · doi:10.1038/349117a0 · Cited by 3,392 · cited 8×
“The fact that Glu-143 links switch I to helix 5 residues, coupled with its interaction with Arg-123 in loop 8 as part of stabilizing the SAK residues at the nucleotide-binding site, explains the strictly conserved nature of Glu-143 across all GTPases.”
BackgroundInfluentialFundamentals of Enzyme Kinetics
Cornish-Bowden, A. J. (1979) · Elsevier eBooks · doi:10.1016/c2013-0-04130-8 · Cited by 2,079
A Comprehensive Survey of Ras Mutations in Cancer
Prior, I. A., Lewis, P. D., & Mattos, C. (2012) · Cancer Research · doi:10.1158/0008-5472.can-11-2612 · Cited by 2,063 · cited 1×
“It is likely that localization and other large-scale biological contexts play a significant role in the selection of particular mutations in the Ras isoforms associated with specific cancers (76), but it is now clear that conformational states at the active site are affected by sequence at the molecular level in a glo…”
BackgroundSelf-citation
Cited by
Sorted by citations
Items 101–133 of 133
Jiang, Y., Jenjob, R., Ryu, D., Shen, Z., & Yang, S. (2025) · Acta Pharmaceutica Sinica B · doi:10.1016/j.apsb.2025.06.024 · Cited by 2
G12 mutations rewire allosteric communication at the Ras–RalGDS interface
Demirbas, E., Jang, H., Kosoglu, K., Nussinov, R., Gürsoy, A., & Keskin, Ö. (2026) · Biophysical Journal · doi:10.1016/j.bpj.2026.01.053 · Cited by 2
An evolutionarily conserved salt bridge stabilizes the active site for GTP hydrolysis in Rho GTPases
Marcus, K., Schwabe, M., Knihtila, R., & Mattos, C. (2026) · Journal of Biological Chemistry · doi:10.1016/j.jbc.2026.111260 · Cited by 2
Self-citationDuhay, V., Tian, M., Kosieradzka, K., Ebner, M., Lo, W.-T., Krauss, M., Sprengel, H.-L., Voss, M., Riechmann, M., Savas, J. N., et al. (2026) · Nature Communications · doi:10.1038/s41467-026-70345-y · Cited by 4
Mozibullah, M., Seyedi, H. A. E., Khatun, M., & Solayman, M. (2024) · Journal of Genetic Engineering and Biotechnology · doi:10.1016/j.jgeb.2024.100378 · Cited by 2
“38 The NRAS gene encodes a small GTPase that acts as a molecular regulator of various cellular activities (including cell proliferation and survival) and has been reported to be associated with various types of cancer, 7 ,8 ,24 .”
BackgroundAljuhani, T. A., Shaik, N. A., Alqawas, R. T., Bokhary, R., Almutadares, M., Mahdi, H. B. A., Alrayes, N. M., El-Harouni, A. A., Elango, R., Banaganapalli, B., et al. (2024) · Frontiers in Pharmacology · doi:10.3389/fphar.2024.1498295 · Cited by 2
Insights into direct KRAS inhibition strategies for cancer treatment
Li, T., Gu, C., Zhou, C., Mao, C., Yang, K., Xu, J., Lu, T., & Chen, J. (2024) · Future Medicinal Chemistry · doi:10.1080/17568919.2024.2424149 · Cited by 3
The structural basis of the oncogenic mutant K-Ras4B homodimersPreprint
Kosoglu, K., Omur, M. E., Jang, H., Nussinov, R., Keskin, Ö., & Gürsoy, A. (2020) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2020.09.07.285783 · Cited by 2
Zhou, J., Fang, Z., Zhang, W., Rao, G.-W., & Zheng, Q. (2025) · Mini-Reviews in Organic Chemistry · doi:10.2174/0118756298391640250731025705 · Cited by 1
Kötting, C., Rudack, T., & Gerwert, K. B. (2025) · Biological Chemistry · doi:10.1515/hsz-2025-0227 · Cited by 1
“However, the frequency of the oncogenic mutations are di ff erent among the isoforms (Johnson et al. 2017) and some manuscript discuss di ff erences due to allosteric interactions (Volmar et al. 2022).”
Advances and Challenges in KRAS Mutation Detection and Clinical Implications
Mirlohi, M. S., Yousefi, T., Razaviyan, J., Nomiri, S., Pishbin, E., Shabani‐Rad, M., Ahmadian, M. R., & Salami, S. (2025) · Cancers · doi:10.3390/cancers18010031 · Cited by 1
“This highlights the importance of understanding these differences when developing targeted cancer therapies [11,12].”
Disulfide tethering reveals cryptic pockets in oncogenic KRAS
Balius, T. E., Dyba, M. D., Kumari, V., Denson, J. L., Yi, M., Sharma, A. K., Prakash, P., Alberico, E. O., Messing, S. A. J., Wall, V. E., et al. (2026) · Communications Chemistry · doi:10.1038/s42004-026-01992-x · Cited by 1
“We speculate that small molecules binding to these pockets may have surprising and varying biological effects, acting through allostery or disruption of effector binding 45,53 .”
Divergent CRD ‐dependent mechanisms govern RAS isoform‐selective recruitment of CRAF and ARAF
Banerjee, S., Malasani, S., Banerjee, S., Vasquez, M. C. L., McSorley, S., & Wang, Z. (2026) · Protein Science · doi:10.1002/pro.70687 · Cited by 1
“Collectively, these results reveal that the ARAF RBD exhibits overall low affinity toward all RAS isoforms, with a clear isoform-selective ranking: KRAS > HRAS > NRAS.”
InfluentialMotif-guided identification of KRAS-interacting proteins
Wu, S., Gao, X., Wu, D., Liu, L., Yao, H., Meng, X., Zhang, X., & Bai, F. (2024) · BMC Biology · doi:10.1186/s12915-024-02067-w · Cited by 1
“Taken BRAF, a positive interaction partner which was verified above, as an example, we applied 3DPPI-Miner to predict the complex structures of BRAF with HRAS and NRAS, two family members of RAS, and evaluate their binding affinities (PDB code: 5J2R, 1CTQ, and 5UHV, respectively) [40–42].”
Oncogenic G12D mutation alters local conformations and dynamics of K-RasPreprint
Vatansever, S., Erman, B., & Gumus, Z. H. (2017) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/178483 · Cited by 1
Combined computational and solution scattering methods to study the dynamical properties of Ras
Guterres (2018) · Digital Repository Service (Northeastern University) · doi:10.17760/d20287341 · Cited by 1
Marcus (2018) · Digital Repository Service (Northeastern University) · doi:10.17760/d20286915 · Cited by 1
Ilter, M., Kaşmer, R., Jalalypour, F., Atılgan, C., Topcu, O., Karakaş, N., & Şensoy, Ö. (2022) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2022.04.24.489309 · Cited by 1
“Therefore, much attention has been focused on the discovery of allosteric sites that can regulate the function of the protein Buhrman et al. (2010); Ostrem et al. (2013); Fetics et al. (2015); Johnson et al. (2017); McCarthy et al. (2019); Khan et al. (2021).”
BackgroundTargeting Cancer from a Structural Biology Perspective
Reid, D. F., & Mattos, C. (2019) · Springer eBooks · doi:10.1007/978-981-32-9816-3_12
Self-citationAllosterism and Drug Discovery
Bell, J. E., & Bell, J. K. (2021) · Wiley eBooks · doi:10.1002/0471266949.bmc299
A novel role for RASSF1A in the regulation of RAS activation.
Stewart, D. H. (2020) · ThinkIR: The University of Louisville's Institutional Repository (University of Louisville) · doi:10.18297/etd/3505
Role of HRas and NRas in murine lung development and neonatal survival
Fuentes‐Mateos, R. (2021) · Gredos (University of Salamanca) · doi:10.14201/gredos.148393
The structural basis of allosteric communication networks of Ras GTPases and their oncogenic mutants
Volmar, A. Y. (2020) · Digital Repository Service (Northeastern University) · doi:10.17760/d20398260
Redox-sensitive small GTPase H-Ras in murine astrocytes, an in vitro study
Zuchegna, C., de Rosa, V., Boscia, F., Porcellini, A., & Messina, S. (2022) · Redox Report · doi:10.1080/13510002.2022.2094109
“enhancement of the Guanine Nucleotide Exchange of these enzymes [24].”
BackgroundFuentes‐Mateos, R., García‐Navas, R., Calzada, N., Santos, E., & Fernández‐Medarde, A. (2022) · Research Square · doi:10.21203/rs.3.rs-2212577/v1
Unraveling the Significance of Mg2+ Dependency and Nucleotide Binding Specificity of H-RAS
Tagad, A., & Patwari, G. N. (2024) · The Journal of Physical Chemistry B · doi:10.1021/acs.jpcb.3c06998
Allosteric interactions in KRas and HRas: studies of KRas/Calmodulin and HRas/Raf-RBD
Ojeda, M. (2023) · Digital Repository Service (Northeastern University) · doi:10.17760/d20621550
Miyan, J., Kumar, N., Moinuddin, Malik, S. A., Akif, U. M., Hameed, R., Riyazuddin, M., Dubey, P., Lal, J., Kumar, A., et al. (2025) · Oncogene · doi:10.1038/s41388-025-03516-8 · Cited by 1
“The Kd for canonical Ras partner viz c-RAF(RAF1) through which majority of Ras dependent pathways are executed is in the nanomolar range (KD for H-RAS/Raf = 94 ± 4 nM; KD for K-RAS/ Raf = 98 ± 2 nM and KD for N-RAS/Raf = 206 ± 8 nM) [50].”
Johnson, C. W., Oladimeji, R., O’Donohue, E., & Haigis, K. M. (2026) · ACS Chemical Biology · doi:10.1021/acschembio.6c00157
Self-citationTichauer, R. E. (2019) · doi:10.70675/a7e2cd0az2fbcz4835z9a85zc98dd4dff6e9
“Not only Ras crystallographic structures present a poor resolution of this region [30, 50, 55, 62, 63] but also, Kraulis et al NMR spectroscopy investigation [36] has confirmed that this region has significant rapid internal motions on the nanosecond time scale.”
BackgroundInfluentialBaladhye, V., & Das, N. (2026) · International Journal of Chemistry and Technology · doi:10.32571/ijct.1801063
Switch dynamics and activity of GTP-bound HRAS affected by R97 mutations at allosteric helix α3
Li, Y., Wang, J., 沈建忠, Longlong, R. E. N., & Chen, J. (2026) · Physical Chemistry Chemical Physics · doi:10.1039/d6cp03485c
The Inhibitory Effect of Manumycin A on Extracellular Vesicles: What Do We Know So Far?
Abramowicz, A. (2026) · Biology of the Cell · doi:10.1111/boc.70081
Metadata
- Authors
- Johnson, C. W., Reid, D., Parker, J. A., Salter, S., Knihtila, R., Kuzmic, P., & Mattos, C.
- Affiliations
- Northeastern University (US)
- Corresponding author
- Mattos, Carla
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2017-08
- Volume
- 292
- Issue
- 31
- Pages
- 12981-12993
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC5546037
- OpenAlex
- W2717068414
- Cited by
- 133
- References
- 84
- Citations by year
- 2026: 12 · 2025: 7 · 2024: 15 · 2023: 10 · 2022: 20 · 2021: 18 · 2020: 14 · 2019: 20 · 2018: 12 · 2017: 5
- FWCI
- 5.69
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Science Foundation (1517295)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Keywords
- Ras GTPase, KRAS, NRAS, H-Ras, Ras isoforms, allosteric regulation, GTP hydrolysis, enzyme kinetics, Ras-binding domain
- MeSH terms
- Allosteric Regulation, Allosteric Site, Enzyme Stability, Guanosine Triphosphate, Humans, Isoenzymes, Ligands, Membrane Proteins, Models, Molecular, Peptide Fragments, Protein Conformation, Recombinant Proteins, Dinucleoside Phosphates, Proto-Oncogene Proteins p21(ras), Point Mutation, Crystallography, X-Ray, Proto-Oncogene Proteins c-raf, Amino Acid Substitution, Catalytic Domain, GTP Phosphohydrolases, Protein Interaction Domains and Motifs, Biocatalysis
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
