Crystallization of a functionally intact Hsc70 chaperone

Jianwen Jiang, Eileen M Lafer, Rui J Sousa

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Hsp70s are essential chaperones with roles in a variety of cellular processes and representatives in all kingdoms of life. They are comprised of a nucleotide-binding domain (NBD) and a protein substrate-binding domain (SBD). Structures of isolated NBDs and SBDs have been reported but, until recently, a functionally intact Hsp70 containing both the NBD and SBD has resisted structure determination. Here, it is reported that preparation of diffraction-quality crystals of functionally intact bovine Hsc70 required (i) deletion of part of the protein to reduce oligomerization, (ii) point mutations in the interface between the SBD and NBD and (iii) use of high concentrations of the structure-stabilizing agents glycerol and trimethylamine oxide (TMAO). The introduction of point mutations in interdomain interfaces and the use of the potent structure stabilizer TMAO may be generally useful in crystallization of multidomain proteins that exhibit interdomain motions.

Original languageEnglish (US)
Pages (from-to)39-43
Number of pages5
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume62
Issue number1
DOIs
StatePublished - Apr 2006

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Crystallization
Nucleotides
crystallization
Point Mutation
nucleotides
Substrates
Oligomerization
Proteins
Excipients
mutations
proteins
Protein Binding
Glycerol
Diffraction
deletion
Crystals
oxides
glycerols
preparation
trimethyloxamine

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Structural Biology
  • Genetics
  • Condensed Matter Physics

Cite this

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