Proximity labeling and orthogonal nanobody pulldown (ID-oPD) approaches to map the spinophilin interactome uncover a putative role for spinophilin in protein homeostasis

dc.contributor.authorClaeboe, Emily T.
dc.contributor.authorBlake, Keyana L.
dc.contributor.authorShah, Nikhil R.
dc.contributor.authorMorris, Cameron W.
dc.contributor.authorHens, Basant
dc.contributor.authorAtwood, Brady K.
dc.contributor.authorAbsalon, Sabrina
dc.contributor.authorMosley, Amber L.
dc.contributor.authorDoud, Emma H.
dc.contributor.authorBaucum, Anthony J., II
dc.contributor.departmentBiochemistry and Molecular Biology, School of Medicine
dc.date.accessioned2025-03-19T08:18:36Z
dc.date.available2025-03-19T08:18:36Z
dc.date.issued2025-01-23
dc.description.abstractSpinophilin is a dendritic spine enriched scaffolding and protein phosphatase 1 targeting protein. To detail spinophilin interacting proteins, we created an Ultra-ID and ALFA-tagged spinophilin encoding construct that permits proximity labeling and orthogonal nanobody pulldown (ID-oPD) of spinophilin-associated protein complexes in heterologous cells. We identified 614 specific, and 312 specific and selective, spinophilin interacting proteins in HEK293 cells and validated a subset of these using orthogonal approaches. Many of these proteins are involved in mRNA processing and translation. In the brain, we determined that spinophilin mRNA is highly neuropil localized and that spinophilin may normally function to limit its own expression but promote the expression of other PSD-associated proteins. Overall, our use of an ID-oPD approach uncovers a novel putative role for spinophilin in mRNA translation and synaptic protein expression specifically within dendritic spines.
dc.eprint.versionPreprint
dc.identifier.citationClaeboe ET, Blake KL, Shah NR, et al. Proximity labeling and orthogonal nanobody pulldown (ID-oPD) approaches to map the spinophilin interactome uncover a putative role for spinophilin in protein homeostasis. Preprint. bioRxiv. 2025;2025.01.23.634546. Published 2025 Jan 23. doi:10.1101/2025.01.23.634546
dc.identifier.urihttps://hdl.handle.net/1805/46353
dc.language.isoen_US
dc.publisherbioRxiv
dc.relation.isversionof10.1101/2025.01.23.634546
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.sourcePMC
dc.subjectSpinophilin
dc.subjectHomeostasis
dc.subjectHeterologous cells
dc.titleProximity labeling and orthogonal nanobody pulldown (ID-oPD) approaches to map the spinophilin interactome uncover a putative role for spinophilin in protein homeostasis
dc.typeArticle
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