title: Structural evaluation of a nanobody targeting complement receptor Vsig4 and its cross reactivity.
abstract: Vsig4 is a recently identified immune regulatory protein related to the B7 family with dual functionality: a negative regulator of T cell activation and a receptor for the complement components C3b and C3c. Here we present a structural evaluation of a nanobody, Nb119, against the extracellular IgV domain protein of both mouse and human recombinant Vsig4, which have a high degree of sequence identity. Although mouse and human Vsig4 bind to Nb119 with a 250 times difference in dissociation constants, the interaction results in a highly identical assembly with a RMSD of 0.4Å. The molecular determinants for Vsig4 recognition and cross reactivity unveiled by the atomic structure of Nb119 in complex with mVsig4 and hVsig4 afford new insights useful for the further optimization of the nanobody for potential use in humans. Additionally, structural analysis of the Vsig4-Nb119 complexes indicates that Nb119 occupies the interface on Vsig4 recognized by the macroglobulin-like domains MG4 and MG5 of C3b. Thus an affinity-improved Nb119 may have the potential to influence the activation of both T cells and complement.
authors.affiliation: Center for Translational Medicine, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, China.
authors.forename: Yurong
authors.initials: Y
authors.lastname: Wen
authors.affiliation: Center for Translational Medicine, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, China.
authors.forename: Zhenlin
authors.initials: Z
authors.lastname: Ouyang
authors.affiliation: Research Group of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium; Laboratory of Myeloid Cell Immunology, VIB Inflammation Research Center, Ghent, Belgium.
authors.forename: Steve
authors.initials: S
authors.lastname: Schoonooghe
authors.affiliation: Department of Biochemistry and Molecular Biology, Key Laboratory of Environment and Genes Related to Diseases, Health Science Center, Xi'an Jiaotong University, Xi'an 710049, China.
authors.forename: Siyu
authors.initials: S
authors.lastname: Luo
authors.affiliation: Research Group of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium; Laboratory of Myeloid Cell Immunology, VIB Inflammation Research Center, Ghent, Belgium.
authors.forename: Patrick
authors.initials: P
authors.lastname: De Baetselier
authors.affiliation: Center for Translational Medicine, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, China.
authors.forename: Wuyuan
authors.initials: W
authors.lastname: Lu
authors.affiliation: Research Group of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium; Laboratory of Myeloid Cell Immunology, VIB Inflammation Research Center, Ghent, Belgium.
authors.forename: Serge
authors.initials: S
authors.lastname: Muyldermans
authors.affiliation: Research Group of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium; Laboratory of Myeloid Cell Immunology, VIB Inflammation Research Center, Ghent, Belgium.
authors.forename: Geert
authors.initials: G
authors.lastname: Raes
authors.affiliation: Department of Biochemistry and Molecular Biology, Key Laboratory of Environment and Genes Related to Diseases, Health Science Center, Xi'an Jiaotong University, Xi'an 710049, China. Electronic address: Fang.Zheng@xjtu.edu.cn.
authors.forename: Fang
authors.initials: F
authors.lastname: Zheng
mesh_terms: D000818:Animals; D000936:Antigen-Antibody Complex / Q000737:chemistry; D002478:Cells, Cultured; D003176:Complement C3 / Q000378:metabolism; D003198:Computer Simulation; D003429:Cross Reactions; D018360:Crystallography, X-Ray; D000939:Epitopes / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D056747:Immunomodulation; D008213:Lymphocyte Activation; D051379:Mice; D018716:Molecular Mimicry; D015394:Molecular Structure; D011485:Protein Binding; D011951:Receptors, Complement / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D061905:Single-Domain Antibodies / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000936:Antigen-Antibody Complex; D003176:Complement C3; D000939:Epitopes; D011951:Receptors, Complement; D061905:Single-Domain Antibodies; C414427:VSIG4 protein, human
keywords: B7 family; CRIg; Complement receptor; Cross reactivity; Crystallography; Nanobody; Vsig4