Crystallization and preliminary X-ray diffraction analysis of a specific VHH domain against mouse prion protein.

TitleCrystallization and preliminary X-ray diffraction analysis of a specific VHH domain against mouse prion protein.
Publication TypeJournal Article
Year of Publication2010
AuthorsAbskharon, R. N. N., S. H. Soror, E. Pardon, H. El Hassan, G. Legname, J. Steyaert, and A. Wohlkonig
JournalActa Crystallogr Sect F Struct Biol Cryst Commun
Volume66
IssuePt 12
Pagination1644-6
Date Published2010 Dec 1
ISSN1744-3091
KeywordsAnimals, Antibodies, Chromatography, Gel, Crystallization, Crystallography, X-Ray, Mice, Prions, Protein Structure, Tertiary, Synchrotrons, X-Ray Diffraction
Abstract

Prion disorders are infectious diseases that are characterized by the conversion of the cellular prion protein PrPC into the pathogenic isoform PrPSc. Specific antibodies that interact with the cellular prion protein have been shown to inhibit this transition. Recombinant VHHs (variable domain of dromedary heavy-chain antibodies) or nanobodies are single-domain antibodies, making them the smallest antigen-binding fragments. A specific nanobody (Nb_PrP_01) was raised against mouse PrPC. A crystallization condition for this recombinant nanobody was identified using high-throughput screening. The crystals were optimized using streak-seeding and the hanging-drop method. The crystals belonged to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a=30.04, b=37.15, c=83.00 Å, and diffracted to 1.23 Å resolution using synchrotron radiation. The crystal structure of this specific nanobody against PrPC together with the known PrPC structure may help in understanding the PrPC/PrPSc transition mechanism.

DOI10.1107/S1744309110042168
Alternate JournalActa Crystallogr. Sect. F Struct. Biol. Cryst. Commun.
PubMed ID21139215
PubMed Central IDPMC2998374