Medium-Scale Structural Genomics: Strategies for Protein Expression and Crystallization - Archive ouverte HAL Access content directly
Journal Articles Accounts of Chemical Research Year : 2003

Medium-Scale Structural Genomics: Strategies for Protein Expression and Crystallization

Renaud Vincentelli
Christophe Bignon
  • Function : Author
  • PersonId : 1092696
Arnaud Gruez
Stéphane Canaan
Mariella Tegoni
  • Function : Author
Rie Campanacci
  • Function : Author
Christian Cambillau
  • Function : Author
  • PersonId : 1092690

Abstract

While high-throughput methods of protein production and crystallization are beginning to be well documented, owing to the output of large structural genomics programs, medium-throughput methods at the laboratory scale lag behind. In this paper, we report a possible way for an academic laboratory to adapt high-throughput to medium-throughput methods, on the basis of the first results of two projects aimed at solving the 3D structures of Escherichia coli and Mycobacterium tuberculosis (Tb) proteins of unknown function. We have developed sequential and iterative procedures as well as new technical processes for these programs. Our results clearly demonstrate the value of this medium-throughput approach. For instance, in the first 14 months of the E. coli program, 69 out of 108 target genes led to soluble proteins, 36 were brought to crystallization, and 28 yielded crystals; among the latter, 13 led to usable data sets and 9 to structures. These results, still incomplete, might help in planning future directions of expression and crystallization of proteins applied to medium-throughput structural genomics programs.

Dates and versions

hal-03160692 , version 1 (05-03-2021)

Identifiers

Cite

Renaud Vincentelli, Christophe Bignon, Arnaud Gruez, Stéphane Canaan, Gerlind Sulzenbacher, et al.. Medium-Scale Structural Genomics: Strategies for Protein Expression and Crystallization. Accounts of Chemical Research, 2003, ⟨10.1021/ar010130s⟩. ⟨hal-03160692⟩
15 View
0 Download

Altmetric

Share

Gmail Facebook X LinkedIn More