Crystallization and preliminary X-ray diffraction analysis of a specific VHH domain against mouse prion protein.
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.
Structural Biology Brussels, Free University of Brussels, Pleinlaan 2, Brussels 1050, Belgium.
Romany N N
RN
Abskharon
Sameh H
SH
Soror
Els
E
Pardon
Hassan
H
El Hassan
Giuseppe
G
Legname
Jan
J
Steyaert
Alexandre
A
Wohlkonig
D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray; D051379:Mice; D000072002:Prion Proteins; D011328:Prions / Q000737:chemistry* / Q000276:immunology*; D017434:Protein Structure, Tertiary; D017356:Synchrotrons; D014961:X-Ray Diffraction
D000906:Antibodies; D000072002:Prion Proteins; D011328:Prions; C512657:Prnp protein, mouse