Crystals of an antibody Fv fragment against an integral membrane protein diffracting to 1.28 A resolution.
The Fv fragment of a monoclonal antibody, 7E2 (IgG1, kappa, murine), which is directed against the integral membrane protein cytochrome c oxidase (EC 1.9.3.1) from Paracoccus denitrificans, was cloned and produced in Escherichia coli. Crystals suitable for high-resolution X-ray analysis were obtained by microdialysis under low salt conditions. The crystals belong to the orthorhombic space group P2(1)2(1)2(1) with unit cell dimensions of a = 51.51 A, b = 56.15 A, c = 99.86 A (1 A = 0.1 nm) and contain one Fv fragment per asymmetric unit. Using synchrotron radiation diffraction data were collected up to 1.28 A resolution. This high resolution is very unusual for a heterodimeric protein. The crystals should open the way for refining not only the atomic positions, but also for obtaining information about internal dynamics.
Ostermeier
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Max-Planck-Institut für Biophysik, Frankfurt/Main, Germany.
Essen
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Michel
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D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D003460:Crystallization; D018076:DNA, Complementary; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000276:immunology*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D010231:Paracoccus denitrificans / Q000737:chemistry / Q000201:enzymology / Q000276:immunology; D010957:Plasmids; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology; D014961:X-Ray Diffraction
D018076:DNA, Complementary; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D003576:Electron Transport Complex IV