FASTDXL: A Generalized Screen to Trap Disulfide-Stabilized Complexes for Use in Structural Studies

Jacob E. Corn, James M. Berger

Research output: Contribution to journalArticle

Abstract

Structural studies of macromolecular complexes have produced extraordinary insights into a wide variety of biological processes. Unfortunately, as structural biologists pursue larger and more challenging assemblies, weakly stable and/or nonspecific interactions can become significant roadblocks to structure determination. We have developed a rapid and effective pool-based screen, termed FASTDXL (focused array screening technique for disulfide X-linking), to produce and identify disulfide-stabilized protein-nucleic acid assemblies. A significant strength of FASTDXL is that it can take advantage of prior structural knowledge about molecular interactions, but does not necessarily rely upon it. A detailed application of the approach to the difficult problem of trapping a bacterial primase-ssDNA complex is described, validating the method as a route toward obtaining diffracting crystals suitable for structure determination.

Original languageEnglish (US)
Pages (from-to)773-780
Number of pages8
JournalStructure
Volume15
Issue number7
DOIs
StatePublished - Jul 18 2007
Externally publishedYes

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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