Open AccessCombining site‐specific mutagenesis and seeding as a strategy to crystallize `difficult' proteins: the case of Staphylococcus aureus thioredoxin
Author(s) -
Roos Goedele,
Brosens Elke,
Wahni Khadija,
Desmyter Aline,
Spinelli Silvia,
Wyns Lode,
Messens Joris,
Loris Remy
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106047075
Subject(s) - thioredoxin , mutant , staphylococcus aureus , mutagenesis , seeding , chemistry , crystallography , biology , biochemistry , genetics , gene , bacteria , agronomy
The P31T mutant of Staphylococcus aureus thioredoxin crystallizes spontaneously in space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 41.7, b = 49.5, c = 55.6 Å. The crystals diffract to 2.2 Å 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.