Structural basis of TMPRSS11D specificity and autocleavage activation DOI Creative Commons
Bryan J. Fraser,

Ryan P. Wilson,

Olzhas Ilyassov

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Окт. 9, 2024

Abstract Transmembrane Protease, Serine-2 (TMPRSS2) and TMPRSS11D are human proteases that enable SARS-CoV-2 Influenza A/B virus infections, but their biochemical mechanisms for facilitating viral cell entry remain unclear. We demonstrate these can spontaneously efficiently cleave own zymogen activation motifs, thereby activating wider protease activity on other cellular substrates. determined co-crystal structures in complexes with a native motif an engineered motif, providing insights into autocleavage revealing unique regions of its substrate binding cleft. further show inhibitor underwent clinical trials TMPRSS2-targeted COVID-19 therapy, nafamostat mesylate, was rapidly cleaved by converted to low derivatives. These tropism liabilities existing serine inhibition strategies will guide future drug discovery campaigns targets.

Язык: Английский

Structural basis of TMPRSS2 zymogen activation and recognition by the HKU1 seasonal coronavirus DOI
I. Fernández, Nell Saunders, S. Duquerroy

и другие.

Cell, Год журнала: 2024, Номер 187(16), С. 4246 - 4260.e16

Опубликована: Авг. 1, 2024

Язык: Английский

Процитировано

14
CRISPR-Cas9 genetic screens reveal regulation of TMPRSS2 by the Elongin BC-VHL complex DOI Creative Commons
Ildar Gabaev,

Alexandra Rowland,

Emilija Jovanovic

и другие.

Scientific Reports, Год журнала: 2025, Номер 15(1)

Опубликована: Апрель 7, 2025

Abstract The TMPRSS2 cell surface protease is used by a broad range of respiratory viruses to facilitate entry into target cells. Together with ACE2, represents key factor for SARS-CoV-2 infection, as mediates cleavage viral spike protein, enabling direct fusion the envelope host membrane. Since start COVID-19 pandemic, has gained attention therapeutic inhibitors which would inhibit but little known about regulation, particularly in types physiologically relevant infection. Here, we performed an unbiased genome-wide CRISPR-Cas9 library screen, together targeted at epigenetic modifiers and transcriptional regulators, identify cellular factors that modulate expression human colon epithelial We find endogenous regulated Elongin BC-VHL complex HIF transcription factors. Depletion B or treatment cells PHD resulted downregulation inhibition show still utilised Omicron variants colonic Our study enhances our understanding regulation

Язык: Английский

Процитировано

0
Structural basis of TMPRSS2 zymogen activation and recognition by the HKU1 seasonal coronavirus DOI Creative Commons
I. Fernández, Nell Saunders, S. Duquerroy

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Фев. 22, 2024

ABSTRACT The human seasonal coronavirus HKU1-CoV, which causes common colds worldwide, relies on the sequential binding to a cell-surface glycan and TMPRSS2 for entry into target cells. is cell surface protease synthesized as zymogen that undergoes autolytic activation process its substrates. Several respiratory viruses - in particular coronaviruses use proteolytic priming of their spike protein drive membrane fusion upon receptor binding. We describe crystal structure HKU1-CoV domain complex with TMPRSS2, showing it recognizes residues lining catalytic groove. Combined mutagenesis interface comparison across species highlight positions 417 469 determinants host tropism. receptor- blocking nanobody or activated further provides structural basis activating conformational change, altering loops recognized by dramatically increasing affinity.

Язык: Английский

Процитировано

2
Engineering customized viral receptors for various coronaviruses DOI Creative Commons
Peng Liu, Meiling Huang, Hua Guo

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Март 4, 2024

Coronaviruses display versatile receptor usage, yet in-depth characterization of coronaviruses lacking known identities has been impeded by the absence feasible infection models. Here, we developed an innovative strategy to engineer functional customized viral receptors (CVRs). The modular design relies on building frameworks comprising various function modules and generating specific epitope-targeting binding domains. We showed key factors for CVRs efficiently facilitate spike cleavage, membrane fusion, pseudovirus entry, authentic virus amplification coronaviruses, resembling their native receptors. Applying this strategy, delineated accessible epitopes SARS-CoV-2 CVR elucidated mechanism entry supported amino-terminus domain (NTD) targeting S2L20-CVR. Furthermore, created CVR-expressing cells assessing antibodies inhibitors against 12 representative from six subgenera, most which Notably, a pan-sarbecovirus sarbecoviruses, as well replicable HKU3 strain RsHuB2019A. Through combining HKU5-specific with reverse genetics, successfully rescued cultured wild-type fluorescence protein-incorporated HKU5, receptor-unidentified merbecovirus. Our study demonstrated great potential in establishing receptor-independent models, paving way studying viruses that are challenging culture due lack susceptible cells.

Язык: Английский

Процитировано

1
Identification of the critical residues of TMPRSS2 for entry and host range of human coronavirus HKU1 DOI
Yahan Chen,

Xiuyuan Ou,

Pei Li

и другие.

Journal of Virology, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 11, 2024

Human coronavirus (CoV) HKU1 infection typically causes common cold but can lead to pneumonia in children, older people, and immunosuppressed individuals. Recently, human transmembrane serine protease 2 (hTMPRSS2) was identified as the functional receptor for HKU1, its region residues critical S binding remain elusive. In this study, we find that could utilize hamster, not rat, mouse, or bat TMPRSS2 virus entry, displaying a narrow host range. Using human-bat chimeras, show peptidase (SP) domain of is essential entry HKU1. Further extensive mutagenesis analyses C-terminal regions SP domains TMPRSS2s identify 417 469 Replacement either D417 Y469 with asparagine hTMPRSS2 abolishes abilities mediate pseudovirions cell-cell fusion, whereas substitution N417 D N469 Y (bTMPRSS2) renders it supporting entry. Our findings contribute deeper understanding coronavirus-receptor interactions cross-species transmission.IMPORTANCEThe proteins their cognate receptors determine range transmission potential. found be Here, bat, serve Moreover, swapping at positions bTMPRSS2 corresponding confers pseudovirions, indicating role these study responsible interaction

Язык: Английский

Процитировано

1
Engineering customized viral receptors for various coronaviruses DOI Creative Commons
Huan Yan, Peng Liu, Meiling Huang

и другие.

Research Square (Research Square), Год журнала: 2024, Номер unknown

Опубликована: Март 25, 2024

Abstract Coronaviruses display versatile receptor usage, yet in-depth characterization of coronaviruses lacking known identities has been impeded by the absence feasible infection models 1,2 . Here, we developed an innovative strategy to engineer functional customized viral receptors (CVRs). The modular design relies on building frameworks comprising various function modules and generating specific epitope-targeting binding domains. We showed key factors for CVRs efficiently facilitate spike cleavage, membrane fusion, pseudovirus entry, authentic virus propagation coronaviruses, resembling their native receptors. Applying this strategy, delineated accessible epitopes SARS-CoV-2 CVR elucidated mechanism entry supported amino-terminus domain (NTD) targeting S2L20-CVR. Furthermore, created CVR-expressing cells assessing antibodies inhibitors against 12 representative from six subgenera, most which Notably, a pan-sarbecovirus sarbecoviruses, as well replicable HKU3 strain RsHuB2019A 3 Through combining HKU5-specific with reverse genetics, successfully rescued cultured wild-type fluorescence protein-incorporated HKU5, receptor-unidentified merbecovirus. Our study demonstrated great potential in establishing receptor-independent models, paving way studying viruses that are challenging culture due lack susceptible cells.

Язык: Английский

Процитировано

0
TMPRSS2 est le récepteur cellulaire du coronavirus saisonnier HKU1 DOI Creative Commons
Nell Saunders, Olivier Schwartz

médecine/sciences, Год журнала: 2024, Номер 40(4), С. 335 - 337

Опубликована: Апрель 1, 2024

Процитировано

0
Structural basis of TMPRSS11D specificity and autocleavage activation DOI Creative Commons
Bryan J. Fraser,

Ryan P. Wilson,

Olzhas Ilyassov

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Окт. 9, 2024

Abstract Transmembrane Protease, Serine-2 (TMPRSS2) and TMPRSS11D are human proteases that enable SARS-CoV-2 Influenza A/B virus infections, but their biochemical mechanisms for facilitating viral cell entry remain unclear. We demonstrate these can spontaneously efficiently cleave own zymogen activation motifs, thereby activating wider protease activity on other cellular substrates. determined co-crystal structures in complexes with a native motif an engineered motif, providing insights into autocleavage revealing unique regions of its substrate binding cleft. further show inhibitor underwent clinical trials TMPRSS2-targeted COVID-19 therapy, nafamostat mesylate, was rapidly cleaved by converted to low derivatives. These tropism liabilities existing serine inhibition strategies will guide future drug discovery campaigns targets.

Язык: Английский

Процитировано

0