Interferon‑induced transmembrane proteins (IFITMs) are frequently overexpressed in cancer cells, including cervical carcinoma cells, and play a role in the progression of various cancer types. However, their mechanisms of action remain incompletely understood. In the present study, by employing a combination of surface membrane protein isolation and quantitative mass spectrometry, it was comprehensively described how the IFITM1 protein influences the composition of the cervical cancer cell surfaceome. Additionally, the effects of interferon‑γ on protein expression and cell surface exposure were evaluated in the presence and absence of IFITM1. The IFITM1‑regulated membrane and membrane‑associated proteins identified are involved mainly in processes such as endocytosis and lysosomal transport, cell‑cell and cell‑extracellular matrix adhesion, antigen presentation and the immune response. To complement the proteomic data, gene expression was analyzed using reverse transcription‑quantitative PCR to distinguish whether the observed changes in protein levels were attributable to transcriptional regulation or differential protein dynamics. Furthermore, the proteomic and gene expression data are supported by functional studies demonstrating the impact of the IFITM1 and IFITM3 proteins on the adhesive, migratory and invasive capabilities of cervical cancer cells, as well as their interactions with immune cells.

• Klíčová slova
• antigen presentation, cervical cancer, immunosurveillance, interferon‑induced transmembrane proteins, invasion, mass spectrometry, membrane protein, migration, surfaceome,
• MeSH
• buněčná adheze MeSH
• diferenciační antigeny * metabolismus   genetika   MeSH
• fenotyp MeSH
• interferon gama farmakologie   metabolismus   MeSH
• lidé MeSH
• membránové proteiny * metabolismus   genetika   MeSH
• nádorové buněčné linie MeSH
• nádory děložního čípku * patologie   genetika   metabolismus   imunologie   MeSH
• pohyb buněk MeSH
• proteiny vázající RNA * metabolismus   genetika   MeSH
• proteom * MeSH
• proteomika metody   MeSH
• regulace genové exprese u nádorů MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• diferenciační antigeny * MeSH
• IFITM3 protein, human MeSH Prohlížeč
• interferon gama MeSH
• leu-13 antigen MeSH Prohlížeč
• membránové proteiny * MeSH
• proteiny vázající RNA * MeSH
• proteom * MeSH

Interferon induced transmembrane proteins (IFITMs) play a dual role in the restriction of RNA viruses and in cancer progression, yet the mechanism of their action remains unknown. Currently, there is no data about the basic biochemical features or biophysical properties of the IFITM1 protein. In this work, we report on description and biochemical characterization of three conformational variants/oligomeric species of recombinant IFITM1 protein derived from an Escherichia coli expression system. The protein was extracted from the membrane fraction, affinity purified, and separated by size exclusion chromatography where two distinct oligomeric species were observed in addition to the expected monomer. These species remained stable upon re-chromatography and were designated as "dimer" and "oligomer" according to their estimated molecular weight. The dimer was found to be less stable compared to the oligomer using circular dichroism thermal denaturation and incubation with a reducing agent. A two-site ELISA and HDX mass spectrometry suggested the existence of structural motif within the N-terminal part of IFITM1 which might be significant in oligomer formation. Together, these data show the unusual propensity of recombinant IFITM1 to naturally assemble into very stable oligomeric species whose study might shed light on IFITM1 anti-viral and pro-oncogenic functions in cells.

• Klíčová slova
• IFITM proteins, hydrogen deuterium exchange, oligomerization, protein conformation, protein structure,
• MeSH
• antivirové látky farmakologie   chemie   metabolismus   MeSH
• diferenciační antigeny * metabolismus   chemie   MeSH
• konformace proteinů * MeSH
• lidé MeSH
• rekombinantní proteiny chemie   izolace a purifikace   metabolismus   biosyntéza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antivirové látky MeSH
• diferenciační antigeny * MeSH
• leu-13 antigen MeSH Prohlížeč
• rekombinantní proteiny MeSH