Quantitative comparison of a human cancer cell surface proteome between interphase and mitosis

• Published in: The EMBO journal 2015-01, Vol.34 (2), p.251-265
• Main Authors: Özlü, Nurhan, Qureshi, Mohammad H, Toyoda, Yusuke, Renard, Bernhard Y, Mollaoglu, Gürkan, Özkan, Nazlı E, Bulbul, Selda, Poser, Ina, Timm, Wiebke, Hyman, Anthony A, Mitchison, Timothy J, Steen, Judith A
• Format: Article
• Language: English
• Subjects: Biomarkers - metabolism; Biotinylation; Cadherins - metabolism; Cancer; Cell cycle; Cell division; cell rounding; cell surface; Chromatography, Affinity; EMBO05; EMBO06; EMBO31; HeLa Cells; Humans; Interphase - physiology; MCF-7 Cells; Membrane Proteins - metabolism; Mitosis - physiology; PCDH7; Proteins; Proteome - analysis; Proteomics - methods; Resource; SILAC; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• ISSN: 0261-4189; 1460-2075
• DOI: 10.15252/embj.201385162

The cell surface is the cellular compartment responsible for communication with the environment. The interior of mammalian cells undergoes dramatic reorganization when cells enter mitosis. These changes are triggered by activation of the CDK1 kinase and have been studied extensively. In contrast, very little is known of the cell surface changes during cell division. We undertook a quantitative proteomic comparison of cell surface‐exposed proteins in human cancer cells that were tightly synchronized in mitosis or interphase. Six hundred and twenty‐eight surface and surface‐associated proteins in HeLa cells were identified; of these, 27 were significantly enriched at the cell surface in mitosis and 37 in interphase. Using imaging techniques, we confirmed the mitosis‐selective cell surface localization of protocadherin PCDH7, a member of a family with anti‐adhesive roles in embryos. We show that PCDH7 is required for development of full mitotic rounding pressure at the onset of mitosis. Our analysis provided basic information on how cell cycle progression affects the cell surface. It also provides potential pharmacodynamic biomarkers for anti‐mitotic cancer chemotherapy. Synopsis Epithelial cells are well known to round up and detach during cell division, but the molecular correlates and mechanisms underlying this process are still unclear. The first quantitative proteomic comparison of HeLa cell surface proteins between interphase and mitosis identifies two anti‐adhesive protocadherins as specific mitotic surface markers with apparent roles in controlling mitotic rounding pressure. SILAC‐LC‐MS/MS identified 628 cell surface‐enriched proteins and quantified their localization during interphase and mitosis. 37 proteins were significantly surface‐enriched in interphase, including proteins known to undergo endocytosis such as transferrin receptor. 27 proteins with significant surface enrichment during mitosis include two protocadherins PCDH1 and PCDH7, whose knockdown leads to decreased cell rounding pressure at the onset of mitosis. PCDH7 undergoes dynamic localization changes from the ER in interphase to the cell surface in mitosis, serving as a selective surface marker that allows enrichment of mitotic cell population. Graphical Abstract Proteomic analysis of cell surface changes during cell division provides insight into molecular correlates of mitotic cell rounding and detachment and identifies mitosis‐selective surface markers with resource potential for a