PT  - JOURNAL ARTICLE
AU  - Daniel Ellis
AU  - Julia Lederhofer
AU  - Oliver J. Acton
AU  - Yaroslav Tsybovsky
AU  - Sally Kephart
AU  - Christina Yap
AU  - Rebecca A. Gillespie
AU  - Adrian Creanga
AU  - Tyler Stephens
AU  - Deleah Pettie
AU  - Michael Murphy
AU  - Andrew J. Borst
AU  - Young-Jun Park
AU  - Kelly K. Lee
AU  - Barney S. Graham
AU  - David Veesler
AU  - Neil P. King
AU  - Masaru Kanekiyo
TI  - Structure-based design of stabilized recombinant influenza neuraminidase tetramers
AID  - 10.1101/2021.05.17.444468
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.05.17.444468
4099  - http://biorxiv.org/content/early/2021/05/17/2021.05.17.444468.short
4100  - http://biorxiv.org/content/early/2021/05/17/2021.05.17.444468.full
AB  - Influenza virus neuraminidase (NA) is a major antiviral drug target and has recently reemerged as a key target of antibody-mediated protective immunity. Here we show that recombinant NAs across all non-bat subtypes adopt various tetrameric conformations, including a previously unreported “open” state that may help explain poorly understood variations in NA stability across viral strains and subtypes. We used homology-directed protein design to uncover the structural principles underlying these distinct tetrameric conformations and stabilize multiple recombinant NAs in the “closed” state. In addition to improving thermal stability, conformational stabilization improved affinity to protective antibodies elicited by viral infection, including antibodies targeting a quaternary epitope and the broadly conserved catalytic site. The stabilized NA proteins can also be integrated into viruses without affecting fitness. Our findings provide a deeper understanding of NA structure, stability, and antigenicity, as well as a roadmap towards structure-based discovery of NA-directed therapeutics and vaccines.Competing Interest StatementD.E., J.L., B.S.G., N.P.K., and M.K. are named as inventors on patent applications filed by the University of Washington based on the studies presented in this paper. N.P.K. is a co-founder, shareholder, paid consultant, and chair of the scientific advisory board of Icosavax, Inc., and has received unrelated sponsored research agreements from Pfizer. D.V. is a consultant for and has received an unrelated sponsored research agreement from Vir Biotechnology, Inc.