Systematic Bioinformatics and Experimental Validation of Yeast Complexes Reduces the Rate of Attrition during Structural Investigations

Mark A. Brooks, Kamil Gewartowski, Eirini Mitsiki, Juliette Letoquart, Roland A. Pache, Ysaline Billier, Michela Bertero, Margot Correa, Mariusz Czarnocki-Cieciura, Michal Dadlez, Veronique Henriot, Noureddine Lazar, Lila Delbos, Dorothee Lebert, Jan Piwowarski, Pascal Rochaix, Bettina Böettcher, Luis Serrano, Bertrand Seraphin, Herman van TilbeurghPatrick Aloy, Anastassis Perrakis, Andrzej Dziembowski

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Pages (from-to)1075-1082
Number of pages8
JournalStructure
Volume18
Issue number9
Early online date7 Sep 2010
DOIs
Publication statusPublished - 8 Sep 2010

Cite this