气候变化领域集成服务门户(CCPortal): A nanobody toolbox targeting dimeric coiled-coil modules for functionalization of designed protein origami structures

CCPortal

DOI	10.1073/pnas.2021899118
	A nanobody toolbox targeting dimeric coiled-coil modules for functionalization of designed protein origami structures
	Majerle A.; Hadzi S.; Aupič J.; Satler T.; Lapenta F.; Strmšek Ž.; Lah J.; Loris R.; Jerala R.
发表日期	2021
ISSN	00278424
卷号	118 期号:17
英文摘要	Coiled-coil (CC) dimers are widely used in protein design because of their modularity and well-understood sequence–structure relationship. In CC protein origami design, a polypeptide chain is assembled from a defined sequence of CC building segments that determine the self-assembly of protein cages into polyhedral shapes, such as the tetrahedron, triangular prism, or four-sided pyramid. However, a targeted functionalization of the CC modules could significantly expand the versatility of protein origami scaffolds. Here, we describe a panel of single-chain camelid antibodies (nanobodies) directed against different CC modules of a de novo designed protein origami tetrahedron. We show that these nanobodies are able to recognize the same CC modules in different polyhedral contexts, such as isolated CC dimers, tetrahedra, triangular prisms, or trigonal bipyramids, thereby extending the ability to functionalize polyhedra with nanobodies in a desired stoichiometry. Crystal structures of five nanobody-CC complexes in combination with small-angle X-ray scattering show binding interactions between nanobodies and CC dimers forming the edges of a tetrahedron with the nanobody entering the tetrahedral cavity. Furthermore, we identified a pair of allosteric nanobodies in which the binding to the distant epitopes on the antiparallel homodimeric APH CC is coupled via a strong positive cooperativity. A toolbox of well-characterized nanobodies specific for CC modules provides a unique tool to target defined sites in the designed protein structures, thus opening numerous opportunities for the functionalization of CC protein origami polyhedra or CC-based bionanomaterials. © 2021 National Academy of Sciences. All rights reserved.
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/179727
作者单位	Department of Synthetic Biology and Immunology, National Institute of Chemistry, Ljubljana, SI-1000, Slovenia; Department of Physical Chemistry, Faculty of Chemistry and Chemical Technology, University of Ljubljana, Ljubljana, SI-1000, Slovenia; Graduate Study Program, Faculty of Chemistry and Chemical Technology, University of Ljubljana, Ljubljana, SI-1000, Slovenia; Vlaams Instituut voor Biotechnologie, Vrije Universiteit Brussel Center for Structural Biology, Vrije University Brussels, Brussels, B-1050, Belgium
推荐引用方式 GB/T 7714	Majerle A.,Hadzi S.,Aupič J.,et al. A nanobody toolbox targeting dimeric coiled-coil modules for functionalization of designed protein origami structures[J],2021,118(17).
APA	Majerle A..,Hadzi S..,Aupič J..,Satler T..,Lapenta F..,...&Jerala R..(2021).A nanobody toolbox targeting dimeric coiled-coil modules for functionalization of designed protein origami structures.Proceedings of the National Academy of Sciences of the United States of America,118(17).
MLA	Majerle A.,et al."A nanobody toolbox targeting dimeric coiled-coil modules for functionalization of designed protein origami structures".Proceedings of the National Academy of Sciences of the United States of America 118.17(2021).