Crystallization and preliminary diffraction studies of prostaglandin E2-specific monoclonal antibody Fab fragment in the ligand complex.
Prostaglandin E(2) is a major lipid mediator that regulates diverse biological processes. To elucidate how prostaglandin E(2) is recognized specifically by its antibody, the Fab fragment of a monoclonal anti-prostaglandin E(2) antibody was prepared and its complex with prostaglandin E(2) was crystallized. The stable Fab-prostaglandin E(2) complex was prepared by gel-filtration chromatography. Crystals were obtained by the microbatch method at 277 K using polyethylene glycol 4000 as a precipitant. A diffraction data set was collected to 2.2 A resolution. The crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 70.3, b = 81.8, c = 82.2 A. The asymmetric unit was suggested to contain one molecule of the Fab-prostaglandin E(2) complex, with a corresponding crystal volume per protein weight of 2.75 A(3) Da(-1).
Kurahashi
Yuko
Y
Faculty of Human Life and Science, Doshisha Women's College of Liberal Arts, Imadegawa-douri Tera-machi Nishi-iru, Kamigyou, Kyoto 602-0893, Japan.
Sugahara
Mitsuaki
M
Ago
Hideo
H
Aoyama
Sae
S
Takahashi
Naoko
N
Takio
Koji
K
Katsukawa
Michiko
M
Yamamoto
Shozo
S
Miyano
Masashi
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D015232:Dinoprostone / Q000737:chemistry / Q000276:immunology; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry* / Q000276:immunology; D008024:Ligands; D051379:Mice; D015394:Molecular Structure; D014961:X-Ray Diffraction
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007140:Immunoglobulin Fab Fragments; D008024:Ligands; D015232:Dinoprostone