Fibronectin type III domains engineered to bind CD40L: cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of two complexes

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2013-09, Vol.69 (9), p.1045-1048
• Main Authors: Oganesyan, Vaheh, Ferguson, Andrew, Grinberg, Luba, Wang, Lin, Phipps, Sandrina, Chacko, Benoy, Drabic, Stacey, Thisted, Thomas, Baca, Manuel
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Binding Sites; CD40 Ligand - chemistry; CD40 Ligand - genetics; CD40 Ligand - isolation & purification; CD40L; Cloning; Combinatorial analysis; Crystal structure; Crystallization Communications; Crystallography, X-Ray; Diffraction; Escherichia coli - genetics; Fibronectin; fibronectin type III domains; Fibronectins - chemistry; Fibronectins - genetics; Gene Expression; Human; Humans; Molecular Sequence Data; Peptide Library; Peptides - chemistry; Peptides - genetics; Peptides - isolation & purification; Protein Binding; Protein Structure, Secondary; Proteins; Purification; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Sequence Alignment
• ISSN: 1744-3091; 1744-3091; 2053-230X
• DOI: 10.1107/S1744309113022847

Tn3 proteins are a novel class of binding molecules based on the third fibronectin type III domain of human tenascin C. Target‐specific Tn3 proteins are selected from combinatorial libraries in which three surface‐exposed loops have been diversified. Here, the cocrystallization of two different Tn3 proteins in complex with CD40L, a therapeutic target for immunological disease, is reported. These crystal structures are the first to be reported of Tn3 proteins and will help to reveal how these engineered molecules achieve specific recognition of a cognate target.