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Glycobiology Advance Access published online on August 3, 2006
Glycobiology, doi:10.1093/glycob/cwl036
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© The Author 2006. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org
Received May 30, 2006
Revised August 1, 2006
Accepted August 2, 2006

Article

Flagellasialin: a novel sulfated {alpha}2,9-linked polysialic acid glycoprotein of sea urchin sperm flagella

Shinji Miyata 1, Chihiro Sato 1, Hironobu Kumita 1, Masaru Toriyama 2, Victor D. Vacquier 3, and Ken Kitajima 1 *

1 Graduate School of Bioagricultural Sciences; Bioscience and Biotechnology Center Nagoya University, Nagoya 464-8601, Japan
2 Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Ohya, Shizuoka 422-8529, Japan
3 Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California, San Diego, La Jolla, California 92093-0202, USA

* To whom correspondence should be addressed.
Ken Kitajima, E-mail: kitajima{at}agr.nagoya-u.ac.jp


  Abstract

A novel {alpha}2,9-linked polysialic acid-containing glycoprotein of sea urchin sperm flagella was identified and named "flagellasialin". Flagellasialin from Hemicentrotus pulcherrimus shows a diverse relative molecular mass on SDS-PAGE of 40-80 kDa. Flagellasialin is a 96-amino acid, threonine-rich, heavily O-glycosylated (80-90% by weight) glycoprotein with a single transmembrane segment at its C-terminus and no apparent cytosolic domain. Of 12 extracellular Thr residues, eight are O-glycosylated and three are non-glycosylated. Flagellasialin is highly expressed in testis, but cannot be detected in ovary. The amino acid sequences of flagellasialin from three sea urchin species (H. pulcherrimus, Strongylocentrotus purpuratus and S. franciscanus) are identical, but some species differences exist in the three core glycan structures to which the sulfated {alpha}2,9-linked polyNeu5Ac chain is linked. Finally, treatment of sperm with a specific antibody against the {alpha}2,9-linked polyNeu5Ac structure results in elevation of intracellular Ca2+ and inhibition of sperm motility and fertilization, implicating flagellasialin as a regulator of these critical processes.

Keywords: fertilization/polysialic acid/sea urchin/sperm motility/flagellasialin.
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