TY - JOUR
T1 - The Human Ganglioside Interactome in Live Cells Revealed Using Clickable Photoaffinity Ganglioside Probes
AU - Zhang, Gao Lan
AU - Porter, Mitchell J.
AU - Awol, Abduselam K.
AU - Orsburn, Benjamin C.
AU - Canner, Samuel W.
AU - Gray, Jeffrey J.
AU - O’Meally, Robert N.
AU - Cole, Robert N.
AU - Schnaar, Ronald L.
N1 - Publisher Copyright:
© 2024 American Chemical Society.
PY - 2024/7/3
Y1 - 2024/7/3
N2 - Gangliosides, sialic acid bearing glycosphingolipids, are components of the outer leaflet of plasma membranes of all vertebrate cells. They contribute to cell regulation by interacting with proteins in their own membranes (cis) or their extracellular milieu (trans). As amphipathic membrane constituents, gangliosides present challenges for identifying their ganglioside protein interactome. To meet these challenges, we synthesized bifunctional clickable photoaffinity gangliosides, delivered them to plasma membranes of cultured cells, then captured and identified their interactomes using proteomic mass spectrometry. Installing probes on ganglioside lipid and glycan moieties, we captured cis and trans ganglioside-protein interactions. Ganglioside interactomes varied with the ganglioside structure, cell type, and site of the probe (lipid or glycan). Gene ontology revealed that gangliosides engage with transmembrane transporters and cell adhesion proteins including integrins, cadherins, and laminins. The approach developed is applicable to other gangliosides and cell types, promising to provide insights into molecular and cellular regulation by gangliosides.
AB - Gangliosides, sialic acid bearing glycosphingolipids, are components of the outer leaflet of plasma membranes of all vertebrate cells. They contribute to cell regulation by interacting with proteins in their own membranes (cis) or their extracellular milieu (trans). As amphipathic membrane constituents, gangliosides present challenges for identifying their ganglioside protein interactome. To meet these challenges, we synthesized bifunctional clickable photoaffinity gangliosides, delivered them to plasma membranes of cultured cells, then captured and identified their interactomes using proteomic mass spectrometry. Installing probes on ganglioside lipid and glycan moieties, we captured cis and trans ganglioside-protein interactions. Ganglioside interactomes varied with the ganglioside structure, cell type, and site of the probe (lipid or glycan). Gene ontology revealed that gangliosides engage with transmembrane transporters and cell adhesion proteins including integrins, cadherins, and laminins. The approach developed is applicable to other gangliosides and cell types, promising to provide insights into molecular and cellular regulation by gangliosides.
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U2 - 10.1021/jacs.4c03196
DO - 10.1021/jacs.4c03196
M3 - Article
C2 - 38887845
AN - SCOPUS:85196736290
SN - 0002-7863
VL - 146
SP - 17801
EP - 17816
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 26
ER -