RESEARCH ARTICLE


Secretion of Legumain Increases in Conditioned Medium from DJ-1-Knockout Cells and in Serum from DJ-1-Knockout Mice



Takuya Yamane1, 2, *, Izumi Kato-Ose3, Tatsuji Sakamoto1, 2, Yoshihisa Nakano1
1 Center for Research and Development Bioresources, Research Organization for University-Community Collaborations, Osaka Prefecture University, Sakai, Osaka599-8570, Japan
2 Department of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Sakai, Osaka599-8531, Japan
3 Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-ku, Sapporo060-0812, Japan


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© 2018 Yamane et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Center for Research and Development Bioresources, Research Organization for University-Community Collaborations, Osaka Prefecture University, Sakai, Osaka 599-8570, Japan; Tel: +81-72-254-9937, (ext. 2962); Fax: +81-72-254-9937; E-mail: tyt29194@osakafu-u.ac.jp


Abstract

Background:

Asparaginyl endopeptidase, also known as legumain (EC 3.4.22.34) shows strong activity in the mouse kidney. Legumain is also highly expressed in tumors. DJ-1/PARK7 is a Parkinson’s disease- and cancer-associated protein. DJ-1 is a coactivator of various transcription factors. Recently, we reported that transcription of the legumain gene is regulated by p53 through DJ-1.

Methods:

We measured the secretion levels of legumain in a conditioned medium of DJ-1 knockout cells and in serum from DJ-1 knockout mice using Western blotting and ELISA. We performed immunocytochemical staining of legumain to examine the localization of legumain in DJ-1-knockout cells.

Results:

We found that the secretion levels of legumain were increased in the conditioned medium of DJ-1-knockout cells and in serum from DJ-1-knockout mice. Dot structures of legumain were also increased in DJ-1-knockout cells.

Conclusion:

The results suggest that legumain secretion from DJ-1-knockout cells and in mice increases through its increased expression and accumulation in membrane-associated vesicles.

Keywords: Legumain, DJ-1, Secretion, p53, Western blotting, Knockout mice, Gene.