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Immunohistochemical study of the membrane skeletal protein, membrane protein palmitoylated 6 (MPP6), in the mouse small intestine

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Abstract

The membrane protein palmitoylated (MPP) family belongs to the membrane-associated guanylate kinase (MAGUK) family. MPP1 interacts with the protein 4.1 family member, 4.1R, as a membrane skeletal protein complex in erythrocytes. We previously described the interaction of another MPP family, MPP6, with 4.1G in the mouse peripheral nervous system. In the present study, the immunolocalization of MPP6 in the mouse small intestine was examined and compared with that of E-cadherin, zonula occludens (ZO)-1, and 4.1B, which we previously investigated in intestinal epithelial cells. The immunolocalization of MPP6 was also assessed in the small intestines of 4.1B-deficient (−/−) mice. In the small intestine, Western blotting revealed that the molecular weight of MPP6 was approximately 55-kDa, and MPP6 was immunostained under the cell membranes in the basolateral portions of almost all epithelial cells from the crypts to the villi. The immunostaining pattern of MPP6 in epithelial cells was similar to that of E-cadherin, but differed from that of ZO-1. In intestinal epithelial cells, the immunostained area of MPP6 was slightly different from that of 4.1B, which was restricted to the intestinal villi. The immunolocalization of MPP6 in small intestinal epithelial cells was similar between 4.1B−/− mice and 4.1B+/+ mice. In the immunoprecipitation study, another MAGUK family protein, calcium/calmodulin-dependent serine protein kinase (CASK), was shown to molecularly interact with MPP6. Thus, we herein showed the immunolocalization and interaction proteins of MPP6 in the mouse small intestine, and also that 4.1B in epithelial cells was not essential for the sorting of MPP6.

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Acknowledgments

This work was partially supported by a Grant from the Japanese Society for the Promotion of Science (KAKEN Number 25460267) to N. Terada.

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    Authors and Affiliations

    1. Division of Health Sciences, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto City, Nagano, 390-8621, Japan

      Akio Kamijo & Nobuo Terada

    2. Department of Anatomy and Molecular Histology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 1110 Shimokato, Chuo City, Yamanashi, 409-3898, Japan

      Yurika Saitoh, Nobuhiko Ohno & Shinichi Ohno

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    1. Akio Kamijo
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    2. Yurika Saitoh
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    3. Nobuhiko Ohno
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    Correspondence to Nobuo Terada.

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    Akio Kamijo and Yurika Saitoh have contributed equally to this work.

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    Kamijo, A., Saitoh, Y., Ohno, N. et al. Immunohistochemical study of the membrane skeletal protein, membrane protein palmitoylated 6 (MPP6), in the mouse small intestine. Histochem Cell Biol 145, 81–92 (2016). https://doi.org/10.1007/s00418-015-1374-7

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