Crystallization and Preliminary X-ray Diffraction Study of an Idiotope-Anti-Idiotope Fv-Fv Complex
| Autor: | Xavier Ysern, Anne Houdusse, Ana Cauerhff, Roberto J. Poljak, Fernando A. Goldbaum, J.-L. Eisele, Roy A. Mariuzza, Barry A. Fields |
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| Rok vydání: | 1994 |
| Předmět: |
Stereochemistry
Molecular Sequence Data Immunoglobulin Variable Region Antigen-Antibody Complex Crystallography X-Ray medicine.disease_cause law.invention Affinity chromatography Structural Biology law medicine Amino Acid Sequence Cloning Molecular Crystallization Immunoglobulin Fragments Molecular Biology Escherichia coli Conserved Sequence chemistry.chemical_classification Genes Immunoglobulin biology Resolution (electron density) Antibodies Monoclonal Idiotopes Antibodies Anti-Idiotypic Amino acid Crystallography Isoelectric point chemistry X-ray crystallography biology.protein Muramidase Isoelectric Focusing |
| Zdroj: | Journal of Molecular Biology. 241:739-743 |
| ISSN: | 0022-2836 |
| Popis: | A complex between the Fv fragment of an anti-hen eggwhite lysozyme antibody (D1.3) and the Fv fragment of an antibody specific for an idiotypic determinant of D1.3 has been crystallized in a form suitable for X-ray diffraction analysis. Both Fv fragments were expressed in soluble form in Escherichia coli and purified by affinity chromatography; diffraction-quality crystals were only obtained following separation of each Fv into distinct isoelectric forms. The crystals belong to space C 2, have unit cell dimensions a = 152·8 A, b = 79·4 A, c = 51·5 A, β = 100·2°, and diffract to better than 2·2 A resolution. The solvent content of the crystals is approximately 60% (v/v) with one Fv-Fv complex in the asymmetric unit. The ability to readily express both components of an antigen-antibody system in bacteria will allow us to rigorously assess the energetic contribution of individual amino acids to complex formation through pairwise mutagenesis of interacting residues. |
| Databáze: | OpenAIRE |
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