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Aksenova VIu Khotin MG Turoverova LV Iudintseva NM Magnusson KÉ Pinaev GP Tentler DG 《Tsitologiia》2012,54(1):25-32
Alpha-actinin 4 (ACTN4) belongs to actin binding proteins of the spectrin superfamily. Structural organisation of actin fibres and focal contacts is considered to be its primary function in a cell. Besides that, nucleocytoplasmic shuffling of ACTN4 and its involvement in nuclear processes were demonstrated. Lately, additional isoforms of ACTN4 resulted from an alternative splicing has been described in various cell types and malignant tumours. In this study, we present investigation of a novel ACTN4 isoform of 80 kDa. The isoform was found in human epidermoid carcinoma cells A431, and it was not detected in human skin fibroblasts, normal human keratinocytes and transformed human embryonic cells HEK293T. Analysis of ACTN4 mRNA in A431 cells showed the presence of a splice variant that lacked the exons 2-8. The deleted exons code two calponin homology domains responsible for ACTN4 binding to F-actin. Intracellular distribution of the described ACTN4 isoform (ACTN4ISO) overexpressed in HEK293T cells differed from that of the full size protein. In the cytoplasm, ACTN4ISO was allocated diffusively with no colocalisation with actin cytoskeleton structures. Intranuclear distribution of ACTN4ISO also differed from that of the full size ACTN4. Nevertheless, immunochemical analysis demonstrated possibility of ACTN4ISO to form heterodimers with the full size protein. Additional investigations of novel isoform interactions with ACTN4 protein partners might clarify its functional features in A431 cells. 相似文献
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L. V. Turoverova M. G. Khotin N. M. Yudintseva K. -E. Magnusson M. I. Blinova G. P. Pinaev D. G. Tentler 《Cell and Tissue Biology》2009,3(5):497-502
The extracellular matrix (ECM) is a highly organized multimolecular structure essential for the vital functions of any organism.
Although much of the data of extracellular matrix components has been accumulated, the isolation of an entire set of these
proteins remains a complex procedure due to the high content of fibrillar proteins and proteoglycans, which form multidomain,
netlike structures. In the study presented, we developed a method for isolating ECM proteins from cell cultures. Human epidermoid
carcinoma cells A431 and fibroblasts obtained from normal and scar human skin were used. We showed that EDTA solution removed
cells from culture plates without destroying the cell membranes. Subsequent treatment of remaining ECM proteins with acetic
acid in order to dissociate collagen fibers significantly improved the fractioning of ECM proteins. The extraction of remaining
proteins from the surface of the culture plate was preformed by a buffer developed based on Laemmli probe buffer. Using this
method, we isolated ECM proteins synthesized by cultured cells, and the extracted proteins were suitable for future analysis
by SDS PAGE and two-dimentional electrophoresis, as well as for identifying individual proteins by mass spectrometry. This
study may allow us to compare assortments of ECM proteins isolated from different sources, and elucidate impact of various
proteins on structure and property of extracellular matrix of investigated cells. 相似文献
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Mikhail Khotin Lidia Turoverova Nikolai Barlev Veronika Viktorija Borutinskaite Alexander Vener Karl-Eric Magnusson Dmitri Tentler 《Biochemical and biophysical research communications》2010,397(2):192-196
Alpha-actinin 4 (ACTN4) is an actin-binding protein. In the cytoplasm, ACTN4 participates in structural organisation of the cytoskeleton via cross-linking of actin filaments. Nuclear localisation of ACTN4 has also been reported, but no clear role in the nucleus has been established. In this report, we describe the identification of proteins associated with ACTN4 in the nucleus. A combination of two-dimensional gel electrophoresis (2D-GE) and MALDI-TOF mass-spectrometry revealed a large number of ACTN4-bound proteins that are involved in various aspects of mRNA processing and transport. The association of ACTN4 with different ribonucleoproteins suggests that a major function of nuclear ACTN4 may be regulation of mRNA metabolism and signaling. 相似文献
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Aleksandrova S. A. Nashchekina Yu. A. Nadezhdin S. V. Vasiliev S. A. Savchenko R. R. Pokrovskaya L. A. Blinova M. I. Mikhailova N. A. Khotin M. G. 《Cell and Tissue Biology》2020,14(6):492-500
Cell and Tissue Biology - The secretome of human mesenchymal stem cells (MSCs) after osteogenic differentiation of FetMSC cells in vitro was obtained to develop tissue engineering technologies for... 相似文献
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