JCB logo
amgmicro.com
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
This Article
Right arrow Full Text (PDF, 4164K)
Right arrow Alert me when this article is cited
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JCB
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kabakoff, B.
Right arrow Articles by Lennarz, W. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kabakoff, B.
Right arrow Articles by Lennarz, W. J.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Facebook   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

The Journal of Cell Biology, Vol 111, 391-400, Copyright © 1990 by The Rockefeller University Press

ARTICLES

Inhibition of glycoprotein processing blocks assembly of spicules during development of the sea urchin embryo

B Kabakoff and WJ Lennarz
Department of Biochemistry and Molecular Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.

Previous studies have implicated an 130-kD glycoprotein containing complex, N-linked oligosaccharide chain(s) in the process of spicule formation in sea urchin embryos. To ascertain whether the processing of high mannose oligosaccharides to complex oligosaccharides is necessary for spiculogenesis, intact embryos and cultures of spicule-forming primary mesenchyme cells were treated with glycoprotein processing inhibitors. In both the embryonic and cell culture systems 1- deoxymannojirimycin (1-MMN) and, to a lesser extent, 1-deoxynojirimycin (1-DNJ) inhibited spicule formation. These inhibitors did not affect gastrulation in whole embryos or filopodial network formation in cell cultures. Swainsonine (SWSN) and castanospermine (CSTP) had no effect in either system. Further analysis revealed the following: (a) 1-MMN entered the embryos and blocked glycoprotein processing in the 24-h period before spicule formation as assessed by a twofold increase in endoglycosidase H sensitivity among newly synthesized glycoproteins upon addition of 1-MMN; (b) 1-MMN did not affect general protein synthesis until after its effects on spicule formation were observed; (c) Immunoblot analysis with an antibody directed towards the polypeptide chain of the 130-kD protein (mAb A3) demonstrated that 1- MMN did not affect the level of the polypeptide that is known to be synthesized just before spicule formation; (d) 1-MMN and 1-DNJ almost completely abolished (greater than 95%) the appearance of mAb 1223 reactive complex oligosaccharide moiety associated with the 130-kD glycoprotein; CSTP and SWSN had much less of an effect on expression of this epitope. These results indicate that the conversion of high mannose oligosaccharides to complex oligosaccharides is required for spiculogenesis in sea urchin embryos and they suggest that the 130-kD protein is one of these essential complex glycoproteins.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Facebook Facebook   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:



  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents