Structure-based design of stabilized recombinant influenza neuraminidase tetramers

Daniel Ellis(University of Washington), Julia Lederhofer(National Institutes of Health), Oliver J. Acton(University of Washington), Yaroslav Tsybovsky(Frederick National Laboratory for Cancer Research), Sally Kephart(University of Washington), Christina Yap(National Institutes of Health), Rebecca A. Gillespie(National Institutes of Health), Adrian Creanga(National Institutes of Health), Audrey Olshefsky(University of Washington), Tyler Stephens(Frederick National Laboratory for Cancer Research), Deleah Pettie(University of Washington), Michael Murphy(University of Washington), Claire Sydeman(University of Washington), Maggie Ahlrichs(University of Washington), Sidney Chan(University of Washington), Andrew J. Borst(University of Washington), Young‐Jun Park(Howard Hughes Medical Institute), Kelly K. Lee(University of Washington), Barney S. Graham(National Institutes of Health), David Veesler(Howard Hughes Medical Institute), Neil P. King(University of Washington), Masaru Kanekiyo(National Institutes of Health)
Nature Communications
April 5, 2022
10.1038/s41467-022-29416-z
Cited by 78Open Access
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Abstract

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 non-bat subtypes adopt various tetrameric conformations, including an "open" state that may help explain poorly understood variations in NA stability across viral strains and subtypes. We use homology-directed protein design to uncover the structural principles underlying these distinct tetrameric conformations and stabilize multiple recombinant NAs in the "closed" state, yielding two near-atomic resolution structures of NA by cryo-EM. In addition to enhancing thermal stability, conformational stabilization improves affinity to protective antibodies elicited by viral infection, including antibodies targeting a quaternary epitope and the broadly conserved catalytic site. Stabilized NAs can also be integrated into viruses without affecting fitness. Our findings provide a deeper understanding of NA structure, stability, and antigenicity, and establish design strategies for reinforcing the conformational integrity of recombinant NA proteins.

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