Crystallization and crystal manipulation of a steric chaperone in complex with its lipase substrate.
Bacterial lipases that are secreted via the type II secretion pathway require a lipase-specific foldase in order to obtain their native and biologically active conformation in the periplasmic space. The lipase-foldase complex from Burkholderia glumae (319 and 333 residues, respectively) was crystallized in two crystal forms. One crystal form belongs to space group P3(1)21 (P3(2)21), with unit-cell parameters a = b = 122.3, c = 98.2 A. A procedure is presented which improved the diffraction of these crystals from approximately 5 to 2.95 A. For the second crystal form, which belonged to space group C2 with unit-cell parameters a = 183.0, b = 75.7, c = 116.6 A, X-ray data were collected to 1.85 A.
Pauwels
Kris
K
Department of Molecular and Cellular Interactions, Vlaams Interuniversitair Instituut voor Biotechnologie (VIB6), Pleinlaan 2, Free University of Brussels, 1050 Brussels, Belgium. krpauwel@vub.ac.be
Loris
Remy
R
Vandenbussche
Guy
G
Ruysschaert
Jean Marie
JM
Wyns
Lode
L
Van Gelder
Patrick
P
D001426:Bacterial Proteins / Q000737:chemistry; D019117:Burkholderia / Q000737:chemistry*; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D008049:Lipase / Q000737:chemistry*; D018832:Molecular Chaperones / Q000737:chemistry*; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D018832:Molecular Chaperones; D008049:Lipase; C504169:lipase foldase