Identification of CD68-Associated Proteins in Myeloid Cells Using Proximity-Dependent Biotinylation
Faculty Mentor
Jason Ashley
Presentation Type
Poster
Start Date
4-14-2026 11:30 AM
End Date
4-14-2026 1:30 PM
Location
PUB NCR
Primary Discipline of Presentation
Biology
Abstract
Understanding protein–protein interactions is essential for elucidating cellular signaling pathways and functional protein networks. In this study, we have designed a strategy to identify protein interaction partners of the macrophage/osteoclast protein CD68 using proximity-dependent biotinylation (BioID). Viral vectors were constructed to deliver N- and C-terminal CD68 fusion constructs linked to the hyperactive biotin ligase ultraID. In addition to the CD68 fusions, we also constructed N- and C-terminal UltraID control constructs that localize to the same cellular compartments as CD68 – specifically, lysosomes, endosomes, and the plasma membrane. Viral-based transduction methods will be used to introduce transgenes into cultured conditionally immortalized myeloid progenitor cells. Upon biotin supplementation, ultraID will catalyze biotinylation of CD68-proximal proteins in living cells, enabling their subsequent detection. Expression and biotinylation activity will be validated via quantitative polymerase chain reaction and Western blot analyses. This work will contribute to a deeper understanding of protein interactions in myeloid lineage cells and highlight the effectiveness of BioID as a proteomic discovery tool.
Recommended Citation
Stepniak, Martyna; Simpson, Justin; David, Victoria; and Gonzalez Naranjo, Valeria, "Identification of CD68-Associated Proteins in Myeloid Cells Using Proximity-Dependent Biotinylation" (2026). 2026 Symposium. 2.
https://dc.ewu.edu/srcw_2026/ps_2026/p2_2026/2
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Identification of CD68-Associated Proteins in Myeloid Cells Using Proximity-Dependent Biotinylation
PUB NCR
Understanding protein–protein interactions is essential for elucidating cellular signaling pathways and functional protein networks. In this study, we have designed a strategy to identify protein interaction partners of the macrophage/osteoclast protein CD68 using proximity-dependent biotinylation (BioID). Viral vectors were constructed to deliver N- and C-terminal CD68 fusion constructs linked to the hyperactive biotin ligase ultraID. In addition to the CD68 fusions, we also constructed N- and C-terminal UltraID control constructs that localize to the same cellular compartments as CD68 – specifically, lysosomes, endosomes, and the plasma membrane. Viral-based transduction methods will be used to introduce transgenes into cultured conditionally immortalized myeloid progenitor cells. Upon biotin supplementation, ultraID will catalyze biotinylation of CD68-proximal proteins in living cells, enabling their subsequent detection. Expression and biotinylation activity will be validated via quantitative polymerase chain reaction and Western blot analyses. This work will contribute to a deeper understanding of protein interactions in myeloid lineage cells and highlight the effectiveness of BioID as a proteomic discovery tool.
