Skip Navigation



Glycobiology Advance Access published online on December 15, 2005
Glycobiology, doi:10.1093/glycob/cwj069
This Article
Right arrow Advance Access manuscript (PDF) Freely available
Right arrowOA All Versions of this Article:
16/4/271    most recent
cwj069v2
cwj069v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Oheda, Y.
Right arrow Articles by Itoh, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Oheda, Y.
Right arrow Articles by Itoh, K.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2005. Published by Oxford University Press. All rights reserved. The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org
Received April 27, 2005
Revised November 22, 2005
Accepted December 9, 2005

Article

Elimination of abnormal sialylglycoproteins in fibroblasts with sialidosis and galactosialidosis by normal gene transfer and enzyme replacement

Yukako Oheda 1, Masaharu Kotani 2, Mai Murata 3, Hitoshi Sakuraba 3, Yoshito Kadota 1, Yutaka Tatano 4, Jun Kuwahara 1, and Kohji Itoh 4 *

1 Department of Medicinal Biotechnology, Graduate School of Pharmaceutical Sciences, Institute for Medicinal Resources, The University of Tokushima, 1-78 Sho-machi, Tokushima 770-8505, Japan
2 Department of Clinical Genetics, The Tokyo Metropolitan Institute of Medical Science, Tokyo Metropolitan Organization for Medical Research, 3-18-22 Honkomagome, Bunkyo, Tokyo 113-8613, Japan
3 Department of Clinical Genetics, The Tokyo Metropolitan Institute of Medical Science, Tokyo Metropolitan Organization for Medical Research, 3-18-22 Honkomagome, Bunkyo, Tokyo 113-8613, Japan; CREST, JST, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan
4 Department of Medicinal Biotechnology, Graduate School of Pharmaceutical Sciences, Institute for Medicinal Resources, The University of Tokushima, 1-78 Sho-machi, Tokushima 770-8505, Japan; CREST, JST, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan

* To whom correspondence should be addressed.
Kohji Itoh, E-mail: kitoh{at}ph.tokushima-u.ac.jp


  Abstract

Sialidosis and galactosialidosis are lysosomal storage diseases caused by genetic defects of lysosomal sialidase (neuraiminidase-1; NEU1) and lysosomal protective protein/cathepsin A (PPCA), respectively, associated with a NEU1 deficiency, excessive accumulation of sialylglycoconjugates and development of progressive neurosomatic manifestations; in addition the latter disorder is accompanied by simultaneous deficiencies of {beta}-galactosidase and cathepsin A. We demonstrated that a few soluble N-glycosylated proteins carrying sialyloligosaccharides sensitive to glycopeptidase F can be specifically detected in cultured fibroblasts from sialidosis and galactosialidosis cases by blotting with a Maackia amurensis (MAM) lectin. We also examined the therapeutic effects of normal gene transfer and enzyme replacement by evaluating the decreases in sialylglycoconjugates accumulated in fibroblasts with these NEU1 deficiencies. The specific N-glycosylated proteins detected on MAM lectin blotting as well as the granular lysosomal fluorescence due to an-avidin-FITC/biotinylated MAM lectin conjugate in sialidosis and galactosialidosis fibroblasts disappeared in parallel with restoration of the intracellular NEU1 activity after transfection of the recombinant NEU1 fused to HA tag sequence and the wild-type PPCA cDNA as well as administration of the recombinant PPCA precursor protein. The detection method for the abnormal sialylglycoproteins in cultured cells involving MAM lectin was demonstrated to be useful for not only biochemical and diagnostic analyses of NEU1 deficiencies but also for therapeutic evaluation of these conditions.

Keywords: galactosialidosis/Maackia amurensis lectin/molecular therapy/sialidosis/sialylglycoprotein.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.