Analysis of the molecular determinants for furin cleavage of the spike protein S1/S2 site in defined strains of the prototype coronavirus murine hepatitis virus (MHV).
Analysis of the molecular determinants for furin cleavage of the spike protein S1/S2 site in defined strains of the prototype coronavirus murine hepatitis virus (MHV).
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DOI：
10.1016/j.virusres.2023.199283
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发表时间：
2024-02
期刊：
影响因子：
5
通讯作者：
Whittaker, Gary R.
中科院分区：
文献类型：
--
作者：
Choi, Annette;Kots, Ekaterina D.;Singleton, Deanndria T.;Weinstein, Harel;Whittaker, Gary R.
关键词：
The S1/S2 domain of the MHV-A59 spike protein is not cleaved by furin, despite a high predictive score Molecular dynamics simulations showed that a His residue in the P2 cleavage position of the S1/S2 site of MHV-A59 spike protein fails to properly orient the sidechain of His194 of the furin catalytic triad to make it incompatible with cleavage initiation The Ser/Thr residue in the P1 position of the S1/S2 site of MHV-2 and MHV-S spike protein distorts the conformation of the furin active site to explain the cleavage pattern for these MHV strains These studies suggest an important process of viral adaptation and evolution within the spike S1/S2 structural loop that informs an understanding of coronavirus infection and pathogenesis We analyzed the spike protein S1/S2 cleavage of selected strains of a prototype coronavirus, mouse hepatitis virus (MHV) by the cellular protease furin, in order to understand the structural requirements underlying the sequence selectivity of the scissile segment. The probability of cleavage of selected MHV strains was first evaluated from furin cleavage scores predicted by the ProP computer software, and then cleavage was measured experimentally with a fluorogenic peptide cleavage assay consisting of S1/S2 peptide mimics and purified furin. We found that in vitro cleavability varied across MHV strains in line with predicted results—but with the notable exception of MHV-A59, which was not cleaved despite a high score predicted for its sequence. Using the known X-Ray structure of furin in complex with a substrate-like inhibitor as an initial structural reference, we carried out molecular dynamics (MD) simulations to learn the modes of binding of the peptides in the furin active site, and the suitability of the complex for initiation of the enzymatic cleavage. We identified the 3D structural requirements of the furin active site configuration that enable bound peptides to undergo cleavage, and the way in which the various strains tested experimentally are fulfilling these requirements. We find that despite some flexibility in the organization of the peptide bound to the active site of the enzyme, the presence of a histidine at P2 of MHV-A59 fails to properly orient the sidechain of His194 of the furin catalytic triad and therefore produces a distortion that renders the peptide/complex structural configuration in the active site incompatible with requirements for cleavage initiation. The Ser/Thr in P1 of MHV-2 and MHV-S has a similar effect of distorting the conformation of the furin active site residues produced by the elimination of the canonical salt-bridge formed by arginine in P1 position. This work informs a study of coronavirus infection and pathogenesis with respect to the function of the viral spike protein, and suggests an important process of viral adaptation and evolution within the spike S1/S2 structural loop.
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DOI：
10.1038/nsb941
发表时间：
2003-07-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Henrich, S;Cameron, A;Than, ME
通讯作者：
Than, ME
影响因子：
3.4
作者：
McGibbon, Robert T.;Beauchamp, Kyle A.;Pande, Vijay S.
通讯作者：
Pande, Vijay S.
影响因子：
5.8
作者：
Lubinski B;Fernandes MHV;Frazier L;Tang T;Daniel S;Diel DG;Jaimes JA;Whittaker GR
通讯作者：
Whittaker GR
影响因子：
5.4
作者：
de Haan, CAM;Stadler, K;Rottier, PJM
通讯作者：
Rottier, PJM
影响因子：
30.3
作者：
Escalera A;Gonzalez-Reiche AS;Aslam S;Mena I;Laporte M;Pearl RL;Fossati A;Rathnasinghe R;Alshammary H;van de Guchte A;Farrugia K;Qin Y;Bouhaddou M;Kehrer T;Zuliani-Alvarez L;Meekins DA;Balaraman V;McDowell C;Richt JA;Bajic G;Sordillo EM;Dejosez M;Zwaka TP;Krogan NJ;Simon V;Albrecht RA;van Bakel H;García-Sastre A;Aydillo T
通讯作者：
Aydillo T