Combining site-specific mutagenesis and seeding as a strategy to crystallize 'difficult' proteins: the case of Staphylococcus aureus thioredoxin

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Dec 1;62(Pt 12):1255-8. doi: 10.1107/S1744309106047075. Epub 2006 Nov 30.

Abstract

The P31T mutant of Staphylococcus aureus thioredoxin crystallizes spontaneously in space group P2(1)2(1)2(1), with unit-cell parameters a = 41.7, b = 49.5, c = 55.6 A. The crystals diffract to 2.2 A resolution. Isomorphous crystals of wild-type thioredoxin as well as of other point mutants only grow when seeded with the P31T mutant. These results suggest seeding as a valuable tool complementing surface engineering for proteins that are hard to crystallize.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallization / methods*
  • Crystallography, X-Ray
  • Mutagenesis, Site-Directed / methods
  • Staphylococcus aureus / chemistry*
  • Staphylococcus aureus / genetics
  • Thioredoxins / chemistry*
  • Thioredoxins / genetics
  • Thioredoxins / isolation & purification

Substances

  • Thioredoxins