Crystallization and molecular-replacement studies of a recombinant antigen-binding fragment complexed with single-stranded DNA.
Anti-DNA antibodies have been implicated in autoimmune diseases and also serve as models for understanding protein-DNA recognition. Crystals of a recombinant antigen-binding fragment (Fab) complexed with dT(5) have been obtained and initial phases have been determined using molecular replacement. The crystals diffract to 2.1 A resolution and occupy space group P6(5)22, with unit-cell parameters a = 171.8, c = 144.6 A; there are two Fabs per asymmetric unit. X-PLORdirect rotation-function calculations followed by Patterson correlation filtering were successful when using a Fab search model; however, they failed when using the individual variable and conserved domains of the Fab as search models. AMoRe successfully identified the correct solution in cases where X-PLOR failed.
Prewitt
S P
SP
Department of Chemistry, University of Missouri-Columbia, Columbia, MO 65211, USA.
Komissarov
A A
AA
Deutscher
S L
SL
Tanner
J J
JJ
D000818:Animals; D000974:Antibodies, Antinuclear / Q000737:chemistry*; D000936:Antigen-Antibody Complex / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000276:immunology*; D007128:Immunoglobulin Fragments / Q000737:chemistry; D051379:Mice; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry
D000974:Antibodies, Antinuclear; D000936:Antigen-Antibody Complex; D004277:DNA, Single-Stranded; D007128:Immunoglobulin Fragments; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; C027903:oligo (dT)