PXD038005

PXD038005 is an original dataset announced via ProteomeXchange.

Title	Mapping of Functional SARS-CoV-2 Receptors in Human Lungs Establishes Differences in Variant Binding and SLC1A5 as a Viral Entry Modulator of hACE2
Description	The COVID-19 pandemic is an infectious disease caused by SARS-CoV-2. The first step of SARS-CoV-2 infection is the recognition of angiotensin-converting enzyme 2 (hACE2) receptors by the receptor-binding domain (RBD) of the viral spike (S) glycoprotein. Although the molecular and structural bases of the SARS-CoV-2-RBD/hACE2 interaction have been thoroughly investigated in vitro, the relationship between hACE2 expression and in vivo infection is less understood. Here, we developed an efficient SARS-CoV-2-RBD binding assay suitable for super resolution microscopy and simultaneous hACE2 immunodetection and mapped the correlation between hACE2 receptor abundance and SARS-CoV-2-RBD binding, both in vitro and in human lung biopsies. Next, we explored the specific proteome of SARS-CoV-2-RBD/hACE2 through a comparative mass spectrometry approach. We found that only a minority of hACE2 positive spots are actually SARS-CoV-2-RBD binding sites, and that the relationship between SARS-CoV-2-RBD binding and hACE2 presence is variable, suggesting the existence of additional factors. Indeed, we found several interactors that are involved in receptor localization and viral entry and characterized one of them: SLC1A5, an amino acid transporter. High-resolution receptor-binding studies showed that co-expression of membrane-bound SLC1A5 with hACE2 predicted SARS-CoV-2 binding and entry better than hACE2 expression alone. Accordingly, SLC1A5 depletion reduces SARS-CoV-2 binding and entry. Notably, the Omicron variant is more efficient in binding hACE2 sites, but equally sensitive to SLC1A5 downregulation. We propose a method for mapping functional SARS-CoV-2 receptors in vivo. We confirm the existence of hACE2 co-factors that may contribute to differential sensitivity of cells to infection.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_07:42:35.084.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	alessandro cuomo
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2022-11-07 09:53:50	ID requested
1	2023-01-09 01:12:37	announced
2	2023-01-25 13:27:16	announced	2023-01-25: Updated project metadata.
⏵ 3	2023-11-14 07:42:37	announced	2023-11-14: Updated project metadata.

Miluzio A, Cuomo A, Cordiglieri C, Donnici L, Pesce E, Bombaci M, Conti M, Fasciani A, Terracciano L, Manganaro L, Toccafondi M, Scagliola A, Oliveto S, Ricciardi S, Grifantini R, De Francesco R, Abrignani S, Manfrini N, Biffo S, Mapping of functional SARS-CoV-2 receptors in human lungs establishes differences in variant binding and SLC1A5 as a viral entry modulator of hACE2. EBioMedicine, 87():104390(2023) [pubmed]

ProteomeXchange project tag: Sars-cov-2, Covid-19

submitter keyword: Spike

Super Resolution microscopy

ACE2 co-factors

RBD

viral binding

viral entry

Nicola Manfrini
contact affiliation	National Institute of Molecular Genetics, "Fondazione Romeo ed Enrica Invernizzi", INGM, 20122 Milan, Italy;
contact email	manfrini@ingm.org
lab head
alessandro cuomo
contact affiliation	IEO
contact email	alessandro.cuomo@ieo.it
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2023/01/PXD038005

PRIDE project URI

• PRIDE
  1. PXD038005
     1. Label: PRIDE project
     2. Name: Mapping of Functional SARS-CoV-2 Receptors in Human Lungs Establishes Differences in Variant Binding and SLC1A5 as a Viral Entry Modulator of hACE2