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Influence of point mutations on PR65 conformational adaptability: Insights from molecular simulations and nanoaperture optical tweezers.
Banerjee, Anupam; Mathew, Samuel; Naqvi, Mohsin M; Yilmaz, Sema Z; Zacharopoulou, Maria; Doruker, Pemra; Kumita, Janet R; Yang, Shang-Hua; Gur, Mert; Itzhaki, Laura S; Gordon, Reuven; Bahar, Ivet.
Afiliación
  • Banerjee A; Laufer Center for Physical and Quantitative Biology, Stony Brook University, Stony Brook, NY 11794, USA.
  • Mathew S; Department of Electrical and Computer Engineering, University of Victoria, Victoria V8P 5C2, Canada.
  • Naqvi MM; Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK.
  • Yilmaz SZ; Department of Mechanical Engineering, Istanbul Technical University, 34437 Istanbul, Turkey.
  • Zacharopoulou M; Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK.
  • Doruker P; Department of Computational and Systems Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
  • Kumita JR; Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK.
  • Yang SH; Department of Electrical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan.
  • Gur M; Department of Mechanical Engineering, Istanbul Technical University, 34437 Istanbul, Turkey.
  • Itzhaki LS; Department of Computational and Systems Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA.
  • Gordon R; Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK.
  • Bahar I; Department of Electrical and Computer Engineering, University of Victoria, Victoria V8P 5C2, Canada.
Sci Adv ; 10(22): eadn2208, 2024 May 31.
Article en En | MEDLINE | ID: mdl-38820156
ABSTRACT
PR65 is the HEAT repeat scaffold subunit of the heterotrimeric protein phosphatase 2A (PP2A) and an archetypal tandem repeat protein. Its conformational mechanics plays a crucial role in PP2A function by opening/closing substrate binding/catalysis interface. Using in silico saturation mutagenesis, we identified PR65 "hinge" residues whose substitutions could alter its conformational adaptability and thereby PP2A function, and selected six mutations that were verified to be expressed and soluble. Molecular simulations and nanoaperture optical tweezers revealed consistent results on the specific effects of the mutations on the structure and dynamics of PR65. Two mutants observed in simulations to stabilize extended/open conformations exhibited higher corner frequencies and lower translational scattering in experiments, indicating a shift toward extended conformations, whereas another displayed the opposite features, confirmed by both simulations and experiments. The study highlights the power of single-molecule nanoaperture-based tweezers integrated with in silico approaches for exploring the effect of mutations on protein structure and dynamics.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Conformación Proteica / Mutación Puntual / Pinzas Ópticas / Simulación de Dinámica Molecular Límite: Humans Idioma: En Revista: Sci Adv Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Conformación Proteica / Mutación Puntual / Pinzas Ópticas / Simulación de Dinámica Molecular Límite: Humans Idioma: En Revista: Sci Adv Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos