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Galectin-1 binds mucin in human trophoblast

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Abstract

Mucins are multifunctional highly glycosylated proteins expressed by the female reproductive tract. Differential expression of MUC1 and MUC15 has been shown in trophoblast. This study was undertaken to establish the distribution of mucin(s) in cytotrophoblast cell cultures using anti-bovine submaxillary mucin (BSM) and to investigate the possibility of MUC1/mucin(s) being a binding partner of trophoblast galectin-1. MUC1 is demonstrated here using immunocytochemistry on isolated cytotrophoblast and the HTR-8/SVneo extravillous trophoblast cell line but detection of additional trophoblast mucins cannot be excluded. Western blot analysis showed similar bands ranging from 30 to >200 kDa with anti-BSM and the well-known mucin antibodies HMFG1 and B72.3. Immunocytochemistry and cell-based ELISA data were found to support that all of the antibodies used are reactive with BSM, suggesting the presence of shared epitopes between BSM and trophoblast mucin(s). Binding of galectin-1 to trophoblast MUC1/mucin(s) was analyzed using a solid-phase assay and co-immunoprecipitation. Recombinant galectin-1 binding to isolated trophoblast mucin in solid-phase assay was sensitive to lactose, a carbohydrate inhibitor of galectin binding. In whole HTR-8/SVneo lysates, ~200 kDa mucin was detected in galectin-1 immunoprecipitates, while endogenous galectin-1 was present in BSM-immunoprecipitates. Furthermore, double fluorescence immunocytochemistry showed overlap of galectin-1 and trophoblast mucins at the plasma membrane of HTR-8/SVneo cells. These results suggest that trophoblast mucin(s) could act as binding partners of galectin-1, in a carbohydrate-dependent manner.

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Acknowledgments

This work was funded through project 173004 of the Ministry of Education, Science and Technological Development, Republic of Serbia.

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Authors and Affiliations

  1. Institute for the Application of Nuclear Energy, INEP, University of Belgrade, Belgrade, Serbia

    Žanka Bojić-Trbojević, Milica Jovanović Krivokuća, Nikola Kolundžić, Snežana Golubović & Ljiljana Vićovac

  2. Faculty of Medicine, Clinic of Gynecology and Obstetrics, Clinical Center of Serbia, University of Belgrade, Belgrade, Serbia

    Miloš Petronijević & Svetlana Vrzić-Petronijević

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  1. Žanka Bojić-Trbojević
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  2. Milica Jovanović Krivokuća
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  3. Nikola Kolundžić
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Correspondence to Žanka Bojić-Trbojević.

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Bojić-Trbojević, Ž., Jovanović Krivokuća, M., Kolundžić, N. et al. Galectin-1 binds mucin in human trophoblast. Histochem Cell Biol 142, 541–553 (2014). https://doi.org/10.1007/s00418-014-1229-7

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