Production, crystallization and X-ray diffraction analysis of two nanobodies against the Duffy binding-like (DBL) domain DBL6ℇ-FCR3 of the Plasmodium falciparum VAR2CSA protein.

TitleProduction, crystallization and X-ray diffraction analysis of two nanobodies against the Duffy binding-like (DBL) domain DBL6ℇ-FCR3 of the Plasmodium falciparum VAR2CSA protein.
Publication TypeJournal Article
Year of Publication2013
AuthorsVuchelen, A., E. Pardon, J. Steyaert, B. Gamain, R. Loris, N. A. J. van Nuland, and S. Ramboarina
JournalActa Crystallogr Sect F Struct Biol Cryst Commun
Volume69
IssuePt 3
Pagination270-4
Date Published2013 Mar 1
ISSN1744-3091
KeywordsAmino Acid Sequence, Animals, Antibodies, Protozoan, Antigens, Protozoan, Binding Sites, Camelids, New World, Crystallography, X-Ray, Erythrocytes, Escherichia coli, Models, Molecular, Molecular Sequence Data, Plasmodium falciparum, Protein Structure, Secondary, Protein Structure, Tertiary, Recombinant Proteins, Sequence Alignment, Sequence Homology, Amino Acid, Single-Domain Antibodies
Abstract

The VAR2CSA protein has been closely associated with pregnancy-associated malaria and is recognized as the main adhesin exposed on the surface of Plasmodium falciparum-infected erythrocytes. Chondroitin sulfate A was identified as the main host receptor in the placenta. Single-domain heavy-chain camelid antibodies, more commonly called nanobodies, were selected and produced against the DBL6ℇ-FCR3 domain of VAR2CSA. Crystals of two specific nanobodies, Nb2907 and Nb2919, identified as strong binders to DBL6ℇ-FCR3 and the full-length VAR2CSA exposed on the surface of FCR3 P. falciparum-infected erythrocytes, were obtained. Crystals of Nb2907 diffract to 2.45 Å resolution and belong to space group C2 with unit-cell parameters a=136.1, b=78.5, c=103.4 Å, β=118.8°, whereas Nb2919 crystals diffract to 2.15 Å resolution and belong to space group P4₃2₁2 with unit-cell parameters a=b=62.7, c=167.2 Å.

DOI10.1107/S1744309113001917
Alternate JournalActa Crystallogr. Sect. F Struct. Biol. Cryst. Commun.
PubMed ID23519802
PubMed Central IDPMC3606572