title: Reconstruction of functional beta-propeller lectins via homo-oligomeric assembly of shorter fragments.
abstract: The modular nature of protein folds suggests that present day proteins evolved via duplication and recombination of smaller functional elements. However, the reconstruction of these putative evolutionary pathways after many millions of years of evolutionary drift has thus far proven difficult, with all attempts to date failing to produce a functional protein. Tachylecin-2 is a monomeric 236 amino acid, five-bladed beta-propeller with five sugar-binding sites. This protein was isolated from a horseshoe crab that emerged ca 500 million years ago. The modular, yet ancient, nature of Tachylectin-2 makes it an excellent model for exploring the evolution of proteins from smaller subunits. To this end, we generated genetically diverse libraries by incremental truncation of the Tachylectin-2 gene and screened them for functional lectins. A number of approximately 100 amino acid residue segments were isolated with the ability to assemble into active homo-pentamers. The topology of most of these segments follows a "hidden" module that differs from the modules observed in wild-type Tachylectin-2, yet their biophysical properties and sugar binding activities resemble the wild-type's. Since the pentamer's molecular mass is twofold higher than the wild-type (approximately 500 amino acid residues), the structure of these oligomeric forms is likely to also differ. Our laboratory evolution experiments highlight the versatility and modularity of the beta-propeller fold, while substantiating the hypothesis that proteins with high internal symmetry, such as beta-propellers, evolved from short, functional gene segments that, at later stages, duplicated, fused, and rearranged, to yield the folds we recognise today.
authors.affiliation: Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Itamar
authors.initials: I
authors.lastname: Yadid
authors.forename: Dan S
authors.initials: DS
authors.lastname: Tawfik
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019020:Directed Molecular Evolution; D019143:Evolution, Molecular; D006737:Horseshoe Crabs / Q000737:chemistry* / Q000378:metabolism; D037102:Lectins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017510:Protein Folding
chemical_list: D037102:Lectins