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Exploring switch II pocket conformation of KRAS(G12D) with mutant-selective monobody inhibitors.
Akkapeddi, Padma; Hattori, Takamitsu; Khan, Imran; Glasser, Eliezra; Koide, Akiko; Ketavarapu, Gayatri; Whaby, Michael; Zuberi, Mariyam; Teng, Kai Wen; Lefler, Julia; Maso, Lorenzo; Bang, Injin; Ostrowski, Michael C; O'Bryan, John P; Koide, Shohei.
Afiliação
  • Akkapeddi P; Laura and Isaac Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016.
  • Hattori T; Laura and Isaac Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016.
  • Khan I; Dertment of Biochemistry and Molecular Pharmacology, New York University Grossman School of Medicine, New York, NY 10016.
  • Glasser E; Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425.
  • Koide A; Ralph H. Johnson VA Medical Center, Charleston, SC 29425.
  • Ketavarapu G; Laura and Isaac Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016.
  • Whaby M; Laura and Isaac Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016.
  • Zuberi M; Department of Medicine, New York University Grossman School of Medicine, New York, NY 10016.
  • Teng KW; Laura and Isaac Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016.
  • Lefler J; Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425.
  • Maso L; Ralph H. Johnson VA Medical Center, Charleston, SC 29425.
  • Bang I; Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425.
  • Ostrowski MC; Ralph H. Johnson VA Medical Center, Charleston, SC 29425.
  • O'Bryan JP; Laura and Isaac Perlmutter Cancer Center, New York University Langone Health, New York, NY 10016.
  • Koide S; Department of Biochemistry and Molecular Biology, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425.
Proc Natl Acad Sci U S A ; 120(28): e2302485120, 2023 07 11.
Article em En | MEDLINE | ID: mdl-37399416
The G12D mutation is among the most common KRAS mutations associated with cancer, in particular, pancreatic cancer. Here, we have developed monobodies, small synthetic binding proteins, that are selective to KRAS(G12D) over KRAS(wild type) and other oncogenic KRAS mutations, as well as over the G12D mutation in HRAS and NRAS. Crystallographic studies revealed that, similar to other KRAS mutant-selective inhibitors, the initial monobody bound to the S-II pocket, the groove between switch II and α3 helix, and captured this pocket in the most widely open form reported to date. Unlike other G12D-selective polypeptides reported to date, the monobody used its backbone NH group to directly recognize the side chain of KRAS Asp12, a feature that closely resembles that of a small-molecule inhibitor, MTRX1133. The monobody also directly interacted with H95, a residue not conserved in RAS isoforms. These features rationalize the high selectivity toward the G12D mutant and the KRAS isoform. Structure-guided affinity maturation resulted in monobodies with low nM KD values. Deep mutational scanning of a monobody generated hundreds of functional and nonfunctional single-point mutants, which identified crucial residues for binding and those that contributed to the selectivity toward the GTP- and GDP-bound states. When expressed in cells as genetically encoded reagents, these monobodies engaged selectively with KRAS(G12D) and inhibited KRAS(G12D)-mediated signaling and tumorigenesis. These results further illustrate the plasticity of the S-II pocket, which may be exploited for the design of next-generation KRAS(G12D)-selective inhibitors.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Neoplasias Pancreáticas / Proteínas Proto-Oncogênicas p21(ras) Idioma: En Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Neoplasias Pancreáticas / Proteínas Proto-Oncogênicas p21(ras) Idioma: En Ano de publicação: 2023 Tipo de documento: Article