Abstract
The Fab fragment of a rat monoclonal antibody (no. 192) with very high affinity for the main immunogenic region of the human muscle nicotinic acetylcholine receptor (AChR) has been purified, characterised and crystallised using vapour diffusion techniques. Its K(d) for human AChR was determined to be 5x10-11 M. Its cross-reactivity pattern suggests that residue α23 of the AChR strongly affects its epitope. Crystals suitable for X-ray analysis, obtained by micro- and macroseeding techniques, belong to the orthorhombic space group C2221 and they diffract to 2.8 Å resolution using synchrotron radiation. The unit cell dimensions are α = 83.4 Å, b = 110.0 Å and c = 212.2 Å and there are two Fab molecules per asymmetric unit.
| Original language | English |
|---|---|
| Pages (from-to) | 195-198 |
| Number of pages | 4 |
| Journal | FEBS Letters |
| Volume | 389 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 1 Jul 1996 |
Funding
Acknowledgements: Supported by grants from the Association Fran-qaise contre les Myopathies, the Biomed-1 program of EU (BMHl-CT93-1100) and the HCM program of EU (CHRXCT94-0547). We are grateful to the staff of the Synchrotron Radiation Source, Dares-bury Laboratory, UK, and the staff of the EMBL outstation at DESY, for help with the data collection, and Drs A. Papageorgiou and D. Leonidas for help with the data processing. We are especially thankful to Prof. M. Kokkinidis, Ms. D. Kotsifaki, Dr. Y. Papani-kolaou and Dr. M. Vlassi (I.M.B.B., Heraclion/Crete) for valuable help with the purification and the crystallisation trials and their general support and encouragement.
Keywords
- Acetylcholine receptor
- Crystallisation
- Main immunogenic region
- Monoclonal antibody
- Myasthenia gravis
- X-ray crystallography
ASJC Scopus subject areas
- Biophysics
- Structural Biology
- Biochemistry
- Molecular Biology
- Genetics
- Cell Biology