Systematic bioinformatics and experimental validation of yeast complexes reduces the rate of attrition during structural investigations - Archive ouverte HAL
Journal Articles Structure (London, England : 1993) Year : 2010

Systematic bioinformatics and experimental validation of yeast complexes reduces the rate of attrition during structural investigations

Mark A. Brooks
  • Function : Author
Kamil Gewartowski
  • Function : Author
Eirini Mitsiki
  • Function : Author
Juliette Létoquart
  • Function : Author
Roland A. Pache
  • Function : Author
Ysaline Billier
  • Function : Author
Michela Bertero
  • Function : Author
Margot Corréa
Mariusz Czarnocki-Cieciura
  • Function : Author
Michal Dadlez
  • Function : Author
Véronique Henriot
  • Function : Author
Noureddine Lazar
  • Function : Author
Lila Delbos
  • Function : Author
Dorothée Lebert
  • Function : Author
Jan Piwowarski
  • Function : Author
Pascal Rochaix
  • Function : Author
Bettina Böttcher
  • Function : Author
Luis Serrano
  • Function : Author
Herman van Tilbeurgh
  • Function : Author
Patrick Aloy
  • Function : Author
Anastassis Perrakis
  • Function : Author
Andrzej Dziembowski
  • Function : Author

Abstract

For high-throughput structural studies of protein complexes of composition inferred from proteomics data, it is crucial that candidate complexes are selected accurately. Herein, we exemplify a procedure that combines a bioinformatics tool for complex selection with in vivo validation, to deliver structural results in a medium-throughout manner. We have selected a set of 20 yeast complexes, which were predicted to be feasible by either an automated bioinformatics algorithm, by manual inspection of primary data, or by literature searches. These complexes were validated with two straightforward and efficient biochemical assays, and heterologous expression technologies of complex components were then used to produce the complexes to assess their feasibility experimentally. Approximately one-half of the selected complexes were useful for structural studies, and we detail one particular success story. Our results underscore the importance of accurate target selection and validation in avoiding transient, unstable, or simply nonexistent complexes from the outset.

Dates and versions

hal-04089578 , version 1 (04-05-2023)

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Cite

Mark A. Brooks, Kamil Gewartowski, Eirini Mitsiki, Juliette Létoquart, Roland A. Pache, et al.. Systematic bioinformatics and experimental validation of yeast complexes reduces the rate of attrition during structural investigations. Structure (London, England : 1993), 2010, 18 (9), pp.1075-1082. ⟨10.1016/j.str.2010.08.001⟩. ⟨hal-04089578⟩
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