title: Connecting two proteins using a fusion alpha helix stabilized by a chemical cross linker.
abstract: Building a sophisticated protein nano-assembly requires a method for linking protein components in a predictable and stable structure. Most of the cross linkers available have flexible spacers. Because of this, the linked hybrids have significant structural flexibility and the relative structure between their two components is largely unpredictable. Here we describe a method of connecting two proteins via a 'fusion α helix' formed by joining two pre-existing helices into a single extended helix. Because simple ligation of two helices does not guarantee the formation of a continuous helix, we used EY-CBS, a synthetic cross linker that has been shown to react selectively with cysteines in α-helices, to stabilize the connecting helix. Formation and stabilization of the fusion helix was confirmed by determining the crystal structures of the fusion proteins with and without bound EY-CBS. Our method should be widely applicable for linking protein building blocks to generate predictable structures.
authors.affiliation: Department of Chemistry, KAIST, Daejeon 34141, Korea.
authors.forename: Woo Hyeon
authors.initials: WH
authors.lastname: Jeong
authors.affiliation: Department of Biological Sciences, KAIST, Daejeon 34141, Korea.
authors.forename: Haerim
authors.initials: H
authors.lastname: Lee
authors.affiliation: Agency for Defense Development, Daejeon 34186, Korea.
authors.forename: Dong Hyun
authors.initials: DH
authors.lastname: Song
authors.affiliation: Department of Chemistry, KAIST, Daejeon 34141, Korea.
authors.forename: Jae-Hoon
authors.initials: JH
authors.lastname: Eom
authors.affiliation: Department of Biological Sciences, KAIST, Daejeon 34141, Korea.
authors.forename: Sun Chang
authors.initials: SC
authors.lastname: Kim
authors.affiliation: Department of Chemistry, KAIST, Daejeon 34141, Korea.
authors.forename: Hee-Seung
authors.initials: HS
authors.lastname: Lee
authors.affiliation: Institute of Biotechnology, Chungnam National University, Daejeon 34134, Korea.
authors.forename: Hayyoung
authors.initials: H
authors.lastname: Lee
authors.affiliation: Department of Chemistry, KAIST, Daejeon 34141, Korea.
authors.forename: Jie-Oh
authors.initials: JO
authors.lastname: Lee
mesh_terms: D017487:Ankyrins / Q000737:chemistry / Q000187:drug effects*; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000737:chemistry / Q000187:drug effects; D010446:Peptide Fragments / Q000737:chemistry / Q000187:drug effects; D017433:Protein Structure, Secondary / Q000187:drug effects; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000187:drug effects*
chemical_list: D017487:Ankyrins; D003432:Cross-Linking Reagents; D010446:Peptide Fragments; D013205:Staphylococcal Protein A; D003545:Cysteine