Crystallization and molecular-replacement studies of the monoclonal antibody mAbR310 specific for the (R)-HNE-modified protein
| Autor: | Kousuke Ishino, Kenichiro Suzuki, Emi Tatsuda, Sohei Ito, Hiroshi Sakai, Koji Uchida |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
medicine.drug_class
Stereochemistry Biophysics Monoclonal antibody Biochemistry law.invention Adduct Immunoglobulin Fab Fragments chemistry.chemical_compound X-Ray Diffraction Structural Biology law Genetics medicine Molecular replacement Crystallization Histidine Aldehydes Chemistry Antibodies Monoclonal Proteins Condensed Matter Physics Papain Crystallography Crystallization Communications Solvents Hemiacetal |
| Zdroj: | Acta Crystallographica Section F Structural Biology and Crystallization Communications. 62:562-564 |
| ISSN: | 1744-3091 |
| DOI: | 10.1107/s1744309106016630 |
| Popis: | 4-hydroxy-2-nonenal (HNE), a major racemic product of lipid peroxidation, reacts with histidine to form a stable HNE-histidine Michael addition-type adduct possessing three chiral centres in the cyclic hemiacetal structure. Monoclonal antibodies against HNE-modified protein have been widely used for assessing oxidative stress in vitro and in vivo. Here, the purification, crystallization and preliminary crystallographic analysis of a Fab fragment of novel monoclonal antibody R310 (mAbR310), which recognizes (R)-HNE-modified protein, are reported. The Fab fragment of mAbR310 was obtained by digestion with papain, purified and crystallized. Using hanging-drop vapour-diffusion crystallization techniques, crystals of mAbR310 Fab were obtained. The crystal belongs to the monoclinic space group C2 (unit-cell parameters a = 127.04, b = 65.31, c = 64.29 A, beta = 118.88 degrees ) and diffracted X-rays to a resolution of 1.84 A. The asymmetric unit contains one molecule of mAbR310, with a corresponding crystal volume per protein weight of 2.51 A(3) Da(-1) and a solvent content of 51.0%. |
| Databáze: | OpenAIRE |
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