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Structure-based Design of Prefusion-stabilized SARS-CoV-2 Spikes
Ching-Lin Hsieh; Jory A Goldsmith; Jeffrey M Schaub; Andrea M DiVenere; Hung-Che Kuo; Kamyab Javanmardi; Kevin C Le; Daniel Wrapp; Alison Gene-Wei Lee; Yutong Liu; Chia-Wei Chou; Patrick O Byrne; Christy K Hjorth; Nicole V Johnson; John Ludes-Meyers; Annalee W Nguyen; Juyeon Park; Nianshuang Wang; Amengor Dzifa; Jennifier A Maynard; Ilya J Finkelstein; Jason S McLellan.
Afiliação
  • Ching-Lin Hsieh; The University of Texas at Austin
  • Jory A Goldsmith; The University of Texas at Austin
  • Jeffrey M Schaub; The University of Texas at Austin
  • Andrea M DiVenere; The University of Texas at Austin
  • Hung-Che Kuo; The University of Texas at Austin
  • Kamyab Javanmardi; The University of Texas at Austin
  • Kevin C Le; The University of Texas at Austin
  • Daniel Wrapp; The University of Texas at Austin
  • Alison Gene-Wei Lee; The University of Texas at Austin
  • Yutong Liu; The University of Texas at Austin
  • Chia-Wei Chou; The University of Texas at Austin
  • Patrick O Byrne; The University of Texas at Austin
  • Christy K Hjorth; The University of Texas at Austin
  • Nicole V Johnson; The University of Texas at Austin
  • John Ludes-Meyers; The University of Texas at Austin
  • Annalee W Nguyen; The University of Texas at Austin
  • Juyeon Park; The University of Texas at Austin
  • Nianshuang Wang; The University of Texas at Austin
  • Amengor Dzifa; The University of Texas at Austin
  • Jennifier A Maynard; The University of Texas at Austin
  • Ilya J Finkelstein; The University of Texas at Austin
  • Jason S McLellan; The University of Texas at Austin
Preprint em Inglês | bioRxiv | ID: ppbiorxiv-125484
ABSTRACT
The COVID-19 pandemic caused by the novel coronavirus SARS-CoV-2 has led to accelerated efforts to develop therapeutics, diagnostics, and vaccines to mitigate this public health emergency. A key target of these efforts is the spike (S) protein, a large trimeric class I fusion protein that is metastable and difficult to produce recombinantly in large quantities. Here, we designed and expressed over 100 structure-guided spike variants based upon a previously determined cryo-EM structure of the prefusion SARS-CoV-2 spike. Biochemical, biophysical and structural characterization of these variants identified numerous individual substitutions that increased protein yields and stability. The best variant, HexaPro, has six beneficial proline substitutions leading to [~]10-fold higher expression than its parental construct and is able to withstand heat stress, storage at room temperature, and multiple freeze-thaws. A 3.2 [A]-resolution cryo-EM structure of HexaPro confirmed that it retains the prefusion spike conformation. High-yield production of a stabilized prefusion spike protein will accelerate the development of vaccines and serological diagnostics for SARS-CoV-2.
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Texto completo: Disponível Coleções: Preprints Base de dados: bioRxiv Idioma: Inglês Ano de publicação: 2020 Tipo de documento: Preprint
Texto completo
Adicionar na Minha BVS
Imprimir
XML
Buscar no Google
Texto completo: Disponível Coleções: Preprints Base de dados: bioRxiv Idioma: Inglês Ano de publicação: 2020 Tipo de documento: Preprint