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Mol. Cells 2010; 30(5): 443-448

Published online November 30, 2010

https://doi.org/10.1007/s10059-010-0138-4

© The Korean Society for Molecular and Cellular Biology

Extracellular Domain of V-Set and Immunoglobulin Domain Containing 1 (VSIG1) Interacts with Ser-toli Cell Membrane Protein, while Its PDZ-Binding Motif Forms a Complex with ZO-1

Ekyune Kim1,4, Youngjeon Lee1,2,4, Ji-Su Kim1, Bong-Seok Song1, Sun-Uk Kim1, Jae-Won Huh1, Sang-Rae Lee1, Sang-Hyun Kim1, Yonggeun Hong2,3, and Kyu-Tae Chang1,*

1National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang 363-883, Korea, 2Department of Rehabilitation Science in Interdisciplinary PhD Program, College of Biomedical Science and Engineering, Inje University, Gimhae 621-749, Korea, 3Department of Physical Therapy, Cardiovascular & Metabolic Disease Center, College of Biomedical Science and Engineering, Inje University, Gimhae 621-749, Korea, 4These authors contributed equally to this work.

Correspondence to : *Correspondence: changkt@kribb.re.kr

Received: May 28, 2010; Revised: August 17, 2010; Accepted: August 19, 2010

Abstract

V-set and immunoglobulin domain containing 1 (VSIG1) is a newly discovered member of the junctional adhesion molecule (JAM) family; it is encoded by a gene located on human chromosome X and preferentially expressed in a variety of cancers in humans. Little is known about its physiological function. To determine the role(s) of VSIG1 in mammalian spermatogenesis, we first generated a specific antibody against mouse VSIG1 and examined the presence and localization of the protein in tissues. RT-RCR and Western blot analysis of the mouse tissues indicated that VSIG1 was specifically expressed in the testis. Furthermore, the results of our trypsinization and biotinylation assays strongly support the assumption that VSIG1 is localized on the testicular germ cell surface. In order to determine whether VSIG1 is capable of participation in homotypic interactions, we performed a GST-pull down assay by using recombinant GST-fusion and His-tagging proteins. The pull-down assay revealed that each GST-fusion Ig-like domain shows homotypic binding. We further show that mVSIG1 can adhere to the Sertoli cells through its first Ig-like domain. To identify the protein that interacted with cytoplasmic domain, we next performed co-immunoprecipitation analysis. This analysis showed that ZO-1, which is the central structural protein of the tight junction, is the binding partner of the cytoplasmic domain of mouse VSIG1. Our findings suggest that mouse VSIG1 interacts with Sertoli cells by heterophilic adhesion via its first Ig-like domain. In addition, its cytoplasmic domain is critical for binding to ZO-1.

Keywords adhesion molecules, JAM family, spermatogenesis, tight junction

Article

Research Article

Mol. Cells 2010; 30(5): 443-448

Published online November 30, 2010 https://doi.org/10.1007/s10059-010-0138-4

Copyright © The Korean Society for Molecular and Cellular Biology.

Extracellular Domain of V-Set and Immunoglobulin Domain Containing 1 (VSIG1) Interacts with Ser-toli Cell Membrane Protein, while Its PDZ-Binding Motif Forms a Complex with ZO-1

Ekyune Kim1,4, Youngjeon Lee1,2,4, Ji-Su Kim1, Bong-Seok Song1, Sun-Uk Kim1, Jae-Won Huh1, Sang-Rae Lee1, Sang-Hyun Kim1, Yonggeun Hong2,3, and Kyu-Tae Chang1,*

1National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Ochang 363-883, Korea, 2Department of Rehabilitation Science in Interdisciplinary PhD Program, College of Biomedical Science and Engineering, Inje University, Gimhae 621-749, Korea, 3Department of Physical Therapy, Cardiovascular & Metabolic Disease Center, College of Biomedical Science and Engineering, Inje University, Gimhae 621-749, Korea, 4These authors contributed equally to this work.

Correspondence to:*Correspondence: changkt@kribb.re.kr

Received: May 28, 2010; Revised: August 17, 2010; Accepted: August 19, 2010

Abstract

V-set and immunoglobulin domain containing 1 (VSIG1) is a newly discovered member of the junctional adhesion molecule (JAM) family; it is encoded by a gene located on human chromosome X and preferentially expressed in a variety of cancers in humans. Little is known about its physiological function. To determine the role(s) of VSIG1 in mammalian spermatogenesis, we first generated a specific antibody against mouse VSIG1 and examined the presence and localization of the protein in tissues. RT-RCR and Western blot analysis of the mouse tissues indicated that VSIG1 was specifically expressed in the testis. Furthermore, the results of our trypsinization and biotinylation assays strongly support the assumption that VSIG1 is localized on the testicular germ cell surface. In order to determine whether VSIG1 is capable of participation in homotypic interactions, we performed a GST-pull down assay by using recombinant GST-fusion and His-tagging proteins. The pull-down assay revealed that each GST-fusion Ig-like domain shows homotypic binding. We further show that mVSIG1 can adhere to the Sertoli cells through its first Ig-like domain. To identify the protein that interacted with cytoplasmic domain, we next performed co-immunoprecipitation analysis. This analysis showed that ZO-1, which is the central structural protein of the tight junction, is the binding partner of the cytoplasmic domain of mouse VSIG1. Our findings suggest that mouse VSIG1 interacts with Sertoli cells by heterophilic adhesion via its first Ig-like domain. In addition, its cytoplasmic domain is critical for binding to ZO-1.

Keywords: adhesion molecules, JAM family, spermatogenesis, tight junction

Mol. Cells
Nov 30, 2022 Vol.45 No.11, pp. 763~867
COVER PICTURE
Naive (cyan) and axotomized (magenta) retinal ganglion cell axons in Xenopus tropicalis (Choi et al., pp. 846-854).

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