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Updated: Feb 15, 2026
Mouse Genome Engineering Using Designer Nucleases
Published on: April 2, 2014
Design of Small Molecules That Compete with Nucleotide Binding to an Engineered Oncogenic KRAS Allele
Yan Zhang, Marie-Hélène Larraufie, Leila Musavi
1Quantitative Proteomics and Metabolomics Center, Columbia University , New York, New York 10027, United States.
Abstract:
RAS mutations are found in 30% of all human cancers, with KRAS the most frequently mutated among the three RAS isoforms (KRAS, NRAS, and HRAS). However, directly targeting oncogenic KRAS with small molecules in the nucleotide-binding site has been difficult because of the high affinity of KRAS for GDP and GTP. We designed an engineered allele of KRAS and a covalent inhibitor that competes for GTP and GDP. This ligand-receptor combination demonstrates that the high affinity of GTP and GDP for RAS proteins can be overcome with a covalent inhibitor and a suitably engineered binding site. The covalent inhibitor irreversibly modifies the protein at the engineered nucleotide-binding site and is able to compete with GDP and GTP. This provides a new tool for studying KRAS function and suggests strategies for targeting the nucleotide-binding site of oncogenic RAS proteins.
Insights
Researchers developed a novel covalent inhibitor and engineered KRAS protein to target cancer-driving mutations. This approach overcomes the high affinity of RAS proteins for GDP and GTP, offering new therapeutic strategies.
Area of Science:
- Oncology
- Molecular Biology
- Drug Discovery
Background:
- RAS mutations, particularly KRAS, are prevalent in 30% of human cancers.
- Targeting oncogenic KRAS is challenging due to its high affinity for GDP and GTP.
Purpose of the Study:
- To design an engineered KRAS allele and a covalent inhibitor to overcome the high affinity of RAS proteins for nucleotides.
- To provide a new tool for studying KRAS function and developing targeted therapies.
Main Methods:
- Engineering a specific KRAS allele with a modified nucleotide-binding site.
- Developing a covalent inhibitor designed to irreversibly bind to the engineered site.
- Demonstrating the inhibitor's ability to compete with GDP and GTP.
Main Results:
- The engineered KRAS-inhibitor system successfully competed with GDP and GTP.
- The covalent inhibitor irreversibly modified the protein at the engineered binding site.
- This demonstrates a viable strategy to overcome the high affinity of RAS proteins for nucleotides.
Conclusions:
- A novel approach using an engineered KRAS allele and covalent inhibitor can overcome the high affinity of RAS proteins for nucleotides.
- This strategy offers a new tool for studying RAS protein function.
- The findings suggest potential therapeutic strategies for targeting oncogenic RAS proteins in cancer.
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