Please use this identifier to cite or link to this item: https://hdl.handle.net/11147/9567
Title: Identification of cytoplasmic sialidase NEU2-associated proteins by LC-MS/MS
Authors: Demir, Seçil Akyıldız
Seyrantepe, Volkan
Demir, Seçil Akyıldız
Seyrantepe, Volkan
Izmir Institute of Technology. Molecular Biology and Genetics
Keywords: Sialidase
NEU2
Actin
Calmodulin
Streptavidin
Issue Date: 2019
Publisher: Türk Biyokimya Derneği
Abstract: Background: Cytoplasmic sialidase (NEU2) plays an active role in removing sialic acids from oligosaccharides, gly-copeptides, and gangliosides in mammalian cells. NEU2 is involved in various cellular events, including cancer metabolism, neuronal and myoblast differentiation, proliferation, and hypertrophy. However, NEU2-interacting protein(s) within the cell have not been identified yet. Objective: The aim of this study is to investigate NEU2 interacting proteins using two-step affinity purification (TAP) strategy combined with mass spectrometry analysis. Methods: In this study, NEU2 gene was cloned into the pCTAP expression vector and transiently transfected to COS-7 cells by using PEI. The most efficient expression time of NEU2- tag protein was determined by real-time PCR and Western blot analysis. NEU2-interacting protein(s) were investigated by using TAP strategy combined with two different mass spectrometry experiment; LC-MS/MS and MALDI TOF/TOF. Results: Here, mass spectrometry analysis showed four proteins; a-actin, beta-actin, calmodulin and histone H1.2 proteins are associated with NEU2. The interactions between NEU2 and actin filaments were verified by Western blot analysis and immunofluorescence analysis. Conclusions: Our study suggests that association of NEU2 with actin filaments and other protein(s) could be important for understanding the biological role of NEU2 in mammalian cells.
Description: WOS: 000489303900006
URI: https://doi.org/10.1515/tjb-2018-0089
https://hdl.handle.net/11147/9567
ISSN: 0250-4685
1303-829X
Appears in Collections:Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection
WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection

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