Systematic bioinformatics and experimental validation of yeast complexes reduces the rate of attrition during structural investigations.
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.
Brooks
Mark A
MA
IBBMC-CNRS UMR8619, IFR 115, Bât. 430, Université Paris-Sud, 91405 Orsay, France.
Gewartowski
Kamil
K
Mitsiki
Eirini
E
Létoquart
Juliette
J
Pache
Roland A
RA
Billier
Ysaline
Y
Bertero
Michela
M
Corréa
Margot
M
Czarnocki-Cieciura
Mariusz
M
Dadlez
Michal
M
Henriot
Véronique
V
Lazar
Noureddine
N
Delbos
Lila
L
Lebert
Dorothée
D
Piwowarski
Jan
J
Rochaix
Pascal
P
Böttcher
Bettina
B
Serrano
Luis
L
Séraphin
Bertrand
B
van Tilbeurgh
Herman
H
Aloy
Patrick
P
Perrakis
Anastassis
A
Dziembowski
Andrzej
A
D019295:Computational Biology / Q000379:methods*; D030562:Databases, Protein; D040901:Proteomics; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism
D029701:Saccharomyces cerevisiae Proteins