tetano
Editor, Senior Moderator
Nat Commun
. 2022 Apr 5;13(1):1825.
doi: 10.1038/s41467-022-29416-z.
Structure-based design of stabilized recombinant influenza neuraminidase tetramers
Daniel Ellis[SUP] #[/SUP][SUP] 1 2 3 4 [/SUP], Julia Lederhofer[SUP] #[/SUP][SUP] 5 [/SUP], Oliver J Acton[SUP] 2 [/SUP], Yaroslav Tsybovsky[SUP] 6 [/SUP], Sally Kephart[SUP] 7 [/SUP], Christina Yap[SUP] 5 [/SUP], Rebecca A Gillespie[SUP] 5 [/SUP], Adrian Creanga[SUP] 5 [/SUP], Audrey Olshefsky[SUP] 1 8 [/SUP], Tyler Stephens[SUP] 6 [/SUP], Deleah Pettie[SUP] 1 2 [/SUP], Michael Murphy[SUP] 1 2 [/SUP], Claire Sydeman[SUP] 1 2 [/SUP], Maggie Ahlrichs[SUP] 1 2 [/SUP], Sidney Chan[SUP] 1 2 [/SUP], Andrew J Borst[SUP] 1 2 [/SUP], Young-Jun Park[SUP] 2 9 [/SUP], Kelly K Lee[SUP] 7 [/SUP], Barney S Graham[SUP] 5 [/SUP], David Veesler[SUP] 2 9 [/SUP], Neil P King[SUP] 10 11 [/SUP], Masaru Kanekiyo[SUP] 12 [/SUP]
Affiliations
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.
. 2022 Apr 5;13(1):1825.
doi: 10.1038/s41467-022-29416-z.
Structure-based design of stabilized recombinant influenza neuraminidase tetramers
Daniel Ellis[SUP] #[/SUP][SUP] 1 2 3 4 [/SUP], Julia Lederhofer[SUP] #[/SUP][SUP] 5 [/SUP], Oliver J Acton[SUP] 2 [/SUP], Yaroslav Tsybovsky[SUP] 6 [/SUP], Sally Kephart[SUP] 7 [/SUP], Christina Yap[SUP] 5 [/SUP], Rebecca A Gillespie[SUP] 5 [/SUP], Adrian Creanga[SUP] 5 [/SUP], Audrey Olshefsky[SUP] 1 8 [/SUP], Tyler Stephens[SUP] 6 [/SUP], Deleah Pettie[SUP] 1 2 [/SUP], Michael Murphy[SUP] 1 2 [/SUP], Claire Sydeman[SUP] 1 2 [/SUP], Maggie Ahlrichs[SUP] 1 2 [/SUP], Sidney Chan[SUP] 1 2 [/SUP], Andrew J Borst[SUP] 1 2 [/SUP], Young-Jun Park[SUP] 2 9 [/SUP], Kelly K Lee[SUP] 7 [/SUP], Barney S Graham[SUP] 5 [/SUP], David Veesler[SUP] 2 9 [/SUP], Neil P King[SUP] 10 11 [/SUP], Masaru Kanekiyo[SUP] 12 [/SUP]
Affiliations
- PMID: 35383176
- DOI: 10.1038/s41467-022-29416-z
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.