Open AccessCrystallization and preliminary X‐ray diffraction analysis of a specific VHH domain against mouse prion protein
Author(s) -
Abskharon Romany N. N.,
Soror Sameh H.,
Pardon Els,
El Hassan Hassan,
Legname Giuseppe,
Steyaert Jan,
Wohlkonig Alexandre
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309110042168
Subject(s) - recombinant dna , prion protein , orthorhombic crystal system , gene isoform , antibody , single domain antibody , crystallization , microbiology and biotechnology , chemistry , virology , crystallography , biophysics , biology , crystal structure , biochemistry , immunology , gene , medicine , disease , organic chemistry , pathology
Prion disorders are infectious diseases that are characterized by the conversion of the cellular prion protein PrP C into the pathogenic isoform PrP Sc . 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 PrP C . 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 P 2 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 PrP C together with the known PrP C structure may help in understanding the PrP C /PrP Sc transition mechanism.