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Structure-function characterization of three human antibodies targeting the vaccinia virus adhesion molecule D8

Matho, Michael H. and Schlossman, Andrew and Gilchuk, Iuliia M. and Miller, Greg and Mikulski, Zbigniew and Hupfer, Matthias and Wang, Jing and Bitra, Aruna and Meng, Xianghzi and Xiang, Yan and Kaever, Tom and Doukov, Tzanko and Ley, Klaus and Crotty, Shane and Peters, Bjoern and Hsieh-Wilson, Linda C. and Crowe, James E., Jr. and Zajonc, Dirk M. (2018) Structure-function characterization of three human antibodies targeting the vaccinia virus adhesion molecule D8. Journal of Biological Chemistry, 293 (1). pp. 390-401. ISSN 0021-9258. PMCID PMC5766908. https://resolver.caltech.edu/CaltechAUTHORS:20171113-124658742

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Abstract

Vaccinia virus (VACV) envelope protein D8 is one of three glycosaminoglycan adhesion molecules and binds to the linear polysaccharide chondroitin sulfate (CS). D8 is also a target for neutralizing antibody responses that are elicited by the smallpox vaccine, which has enabled the first eradication of a human viral pathogen and is a useful model for studying antibody responses. However, to date, VACV epitopes targeted by human antibodies have not been characterized at atomic resolution. Here, we characterized the binding properties of several human anti-D8 antibodies and determined the crystal structures of three VACV-mAb variants, VACV-66, VACV-138, and VACV-304, separately bound to D8. Although all these antibodies bound D8 with high affinity and were moderately neutralizing in the presence of complement, VACV-138 and VACV-304 also fully blocked D8 binding to CS-A, the low affinity ligand for D8. VACV-138 also abrogated D8 binding to the high-affinity ligand CS-E, but we observed residual CS-E binding was observed in the presence of VACV-304. Analysis of the VACV-138– and VACV-304–binding sites along the CS-binding crevice of D8, combined with different efficiencies of blocking D8 adhesion to CS-A and CS-E allowed us to propose that D8 has a high- and low-affinity CS-binding region within its central crevice. The crevice is amenable to protein engineering to further enhance both specificity and affinity of binding to CS-E. Finally, a wild-type D8 tetramer specifically bound to structures within the developing glomeruli of the kidney, which express CS-E. We propose that through structure-based protein engineering, an improved D8 tetramer could be used as a potential diagnostic tool to detect expression of CS-E, which is a possible biomarker for ovarian cancer.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1074/jbc.M117.814541DOIArticle
http://www.ncbi.nlm.nih.gov/pmc/articles/pmc5766908/PubMed CentralArticle
http://www.jbc.org/content/293/1/390/suppl/DC1PublisherSupporting Information
ORCID:
AuthorORCID
Wang, Jing0000-0003-2078-137X
Hsieh-Wilson, Linda C.0000-0001-5661-1714
Crowe, James E., Jr.0000-0002-0049-1079
Zajonc, Dirk M.0000-0001-7215-1981
Additional Information:© 2018 The American Society for Biochemistry and Molecular Biology. Received August 25, 2017; Accepted November 9, 2017; First Published on November 9, 2017. We thank Angela Lamberth for technical assistance in preparing and staining the tissue sections for immunofluorescence microscopy. Use of the Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, is supported by the United States Dept. of Energy, Office of Science, Office of Basic Energy Sciences under Contract No. DE-AC02-76SF00515. The SSRL Structural Molecular Biology Program is supported by the DOE Office of Biological and Environmental Research, and by the National Institutes of Health, National Institute of General Medical Sciences Grant P41GM103393. This work was supported by NIAID, National Institutes of Health, Department of Health and Human Services Contracts HHSN272200900048C (BP), HHSN272200900047C, and HHSN272201400024C (to J. E. C.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The atomic coordinates and structure factors (codes 5USH, 6B9J, and 5USL) have been deposited in the Protein Data Bank (http://wwpdb.org/).
Funders:
Funding AgencyGrant Number
Department of Energy (DOE)DE-AC02-76SF00515
NIHP41GM103393
NIHHHSN272200900048C
NIHHHSN272200900047C
NIHHHSN272201400024C
Subject Keywords:antibody, vaccine, viral protein, glycosaminoglycan, protein crystallization, protein structure, X-ray crystallography
Issue or Number:1
PubMed Central ID:PMC5766908
Record Number:CaltechAUTHORS:20171113-124658742
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20171113-124658742
Official Citation:Michael H. Matho, Andrew Schlossman, Iuliia M. Gilchuk, Greg Miller, Zbigniew Mikulski, Matthias Hupfer, Jing Wang, Aruna Bitra, Xiangzhi Meng, Yan Xiang, Tom Kaever, Tzanko Doukov, Klaus Ley, Shane Crotty, Bjoern Peters, Linda C. Hsieh-Wilson, James E. Crowe Jr., and Dirk M. Zajonc Structure–function characterization of three human antibodies targeting the vaccinia virus adhesion molecule D8 J. Biol. Chem. 2018 293: 390-. doi:10.1074/jbc.M117.814541
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83152
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:14 Nov 2017 22:00
Last Modified:10 Apr 2020 22:07

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