Crystallization and X-ray diffraction data of the cleaved form of plasminogen activator inhibitor-1.
To characterize the structural requirements for the conformational flexibility in plasminogen activator inhibitor-1 (PAI-1) we have crystallized human PAI-1, carrying a mutation which stabilizes PAI-1 in its substrate form. Crystallization was performed by the hanging drop diffusion method at pH 8.5 in the presence of 19% (w/v) polyethyleneglycol 4000 as a precipitant. The crystals appear after 3 days at 23 degrees C and belong to the monoclinic space group C2 with cell dimensions of a = 151.8 A, b = 47.5 A, c = 62.7 A, and beta = 113.9 degrees, and one molecule in the asymmetric unit. The X-ray diffraction data set contains data with a limiting resolution of 2.5 A. Biochemical analysis of the redissolved crystals indicated that during the crystallization process, cleavage had occurred in the active site loop at the P1-P1' position. The availability of good-quality crystals of the cleaved form of this serpin will allow its three-dimensional structure to be solved and will provide detailed information on the structure-function relationship in PAI-1.
Laboratory for Analytical Chemistry and Medicinal Physicochemistry, Faculty of Pharmaceutical Sciences, Katholieke Universiteit Leuven, Belgium.
K
K
Aertgeerts
H L
HL
De Bondt
C
C
De Ranter
P J
PJ
Declerck
D003460:Crystallization; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D017395:Plasminogen Activator Inhibitor 1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
D017395:Plasminogen Activator Inhibitor 1; D011994:Recombinant Proteins