title: A Rhizavidin Monomer with Nearly Multimeric Avidin-Like Binding Stability Against Biotin Conjugates.
abstract: Developing a monomeric form of an avidin-like protein with highly stable biotin binding properties has been a major challenge in biotin-avidin linking technology. Here we report a monomeric avidin-like protein-enhanced monoavidin-with off-rates almost comparable to those of multimeric avidin proteins against various biotin conjugates. Enhanced monoavidin (eMA) was developed from naturally dimeric rhizavidin by optimally maintaining protein rigidity during monomerization and additionally shielding the bound biotin by diverse engineering of the surface residues. eMA allowed the monovalent and nonperturbing labeling of head-group-biotinylated lipids in bilayer membranes. In addition, we fabricated an unprecedented 24-meric avidin probe by fusing eMA to a multimeric cage protein. The 24-meric avidin and eMA were utilized to demonstrate how artificial clustering of cell-surface proteins greatly enhances the internalization rates of assembled proteins on live cells.
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
authors.forename: Jeong Min
authors.initials: JM
authors.lastname: Lee
authors.affiliation: Graduate School of Nanoscience and Technology, Korea Advanced Institute of Science and Technology, Korea.
authors.forename: Jung A
authors.initials: JA
authors.lastname: Kim
authors.affiliation: Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, 10617, Taiwan.
authors.forename: Tzu-Chi
authors.initials: TC
authors.lastname: Yen
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
authors.forename: In Hwan
authors.initials: IH
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
authors.forename: Byungjun
authors.initials: B
authors.lastname: Ahn
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea.
authors.forename: Younghoon
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, 10617, Taiwan.
authors.forename: Chia-Lung
authors.initials: CL
authors.lastname: Hsieh
authors.affiliation: Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Korea.
authors.forename: Ho Min
authors.initials: HM
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Korea Advanced Institute of Science and Technology, Daejeon, 305-701, Korea. ywjung@kaist.ac.kr.
authors.forename: Yongwon
authors.initials: Y
authors.lastname: Jung
mesh_terms: D001360:Avidin / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D001704:Biopolymers / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry*; D011485:Protein Binding
chemical_list: D001426:Bacterial Proteins; D001704:Biopolymers; C535197:rhizavidin, Rhizobium etli; D001360:Avidin; D001710:Biotin
keywords: biotin label; monomeric avidin; multivalency; protein engineering; self-assembly