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10.1371/journal.ppat.1009282

http://scihub22266oqcxt.onion/10.1371/journal.ppat.1009282
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33556147!7895411!33556147
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suck abstract from ncbi


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pmid33556147      PLoS+Pathog 2021 ; 17 (2): e1009282
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  • Multimerization- and glycosylation-dependent receptor binding of SARS-CoV-2 spike proteins #MMPMID33556147
  • Bouwman KM; Tomris I; Turner HL; van der Woude R; Shamorkina TM; Bosman GP; Rockx B; Herfst S; Snijder J; Haagmans BL; Ward AB; Boons GJ; de Vries RP
  • PLoS Pathog 2021[Feb]; 17 (2): e1009282 PMID33556147show ga
  • Receptor binding studies on sarbecoviruses would benefit from an available toolkit of recombinant spike proteins, or domains thereof, that recapitulate receptor binding properties of native viruses. We hypothesized that trimeric Receptor Binding Domain (RBD) proteins would be suitable candidates to study receptor binding properties of SARS-CoV-1 and -2. Here we created monomeric and trimeric fluorescent RBD proteins, derived from adherent HEK293T, as well as in GnTI-/- mutant cells, to analyze the effect of complex vs high mannose glycosylation on receptor binding. The results demonstrate that trimeric, complex glycosylated proteins are superior in receptor binding compared to monomeric and immaturely glycosylated variants. Although differences in binding to commonly used cell lines were minimal between the different RBD preparations, substantial differences were observed when respiratory tissues of experimental animals were stained. The RBD trimers demonstrated distinct ACE2 expression profiles in bronchiolar ducts and confirmed the higher binding affinity of SARS-CoV-2 over SARS-CoV-1. Our results show that complex glycosylated trimeric RBD proteins are attractive to analyze sarbecovirus receptor binding and explore ACE2 expression profiles in tissues.
  • |*Protein Multimerization[MESH]
  • |A549 Cells[MESH]
  • |Angiotensin-Converting Enzyme 2/genetics/*metabolism[MESH]
  • |Animals[MESH]
  • |Chlorocebus aethiops[MESH]
  • |Dogs[MESH]
  • |Glycosylation[MESH]
  • |HEK293 Cells[MESH]
  • |Humans[MESH]
  • |Madin Darby Canine Kidney Cells[MESH]
  • |Mesocricetus[MESH]
  • |Mice[MESH]
  • |N-Acetylglucosaminyltransferases/genetics/metabolism[MESH]
  • |Protein Binding[MESH]
  • |SARS-CoV-2/genetics/*metabolism[MESH]
  • |Spike Glycoprotein, Coronavirus/genetics/*metabolism[MESH]


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