|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Papers In Press, published online ahead of print December 8, 2005
Department für Chemie, Universität für Bodenkultur, Wien 1190
Corresponding Author: renaud.leonard{at}boku.ac.at
Most processed, e.g., fucosylated, N-glycans on insect glycoproteins terminate in mannose, yet the relevant modifying enzymes require the prior action of N-acetylglucos-aminyltransferase I. This led to the hypothesis that a hexosaminidase acts during the course of N-glycan maturation. In order to determine whether the Drosophila melanogaster genome indeed encodes such an enzyme, a cDNA corresponding to fused lobes (fdl), a putative
J. Biol. Chem, 10.1074/jbc.M511023200
Submitted on October 11, 2005
Revised on November 28, 2005
Accepted on December 8, 2005
The Drosophila fused lobes gene encodes an N-acetylglucosaminidase involved in N-glycan processing
-N-acetylglucosaminidase with a potential transmembrane domain, was cloned. When expressed in Pichia pastoris, the enzyme exhibited a substrate specificity similar to that previously described for a hexosaminidase activity from Sf-9 cells, i.e., it hydrolysed exclusively the GlcNAc residue attached to the 
1,3-linked mannose of the core pentasaccharide of N-glycans. It also hydrolysed p-nitrophenyl-N-acetyl--glucosaminide, but not chitooligo-saccharides; in contrast, Drosophila HEXO1 and HEXO2 expressed in Pichia cleaved both these substrates but not N-glycans. The localisation of recombinant FDL tagged with GFP in Drosophila S2 cells by immunoelectron microscopy showed that this enzyme transits through the Golgi, is present on the plasma membrane and in multivesicular bodies and is secreted. Finally, the N-glycans of two lines of fdl mutant flies were analysed by mass spectrometry and reverse phase HPLC. The ratio of structures with terminal GlcNAc over those without (i.e., paucimannosidic N-glycans) was drastically increased in the fdl deficient flies. Therefore, we conclude that the fdl gene encodes a novel hexosaminidase responsible for the occurrence of paucimannosidic N-glycans in Drosophila.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  K. G. T. Hagen, L. Zhang, E Tian, and Y. Zhang Glycobiology on the fly: Developmental and mechanistic insights from Drosophila Glycobiology, February 1, 2009; 19(2): 102 - 111. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. S Lau and J. W Dennis N-Glycans in cancer progression Glycobiology, October 1, 2008; 18(10): 750 - 760. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Geisler, J. J. Aumiller, and D. L. Jarvis A fused lobes Gene Encodes the Processing {beta}-N-Acetylglucosaminidase in Sf9 Cells J. Biol. Chem., April 25, 2008; 283(17): 11330 - 11339. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Koles, J.-M. Lim, K. Aoki, M. Porterfield, M. Tiemeyer, L. Wells, and V. Panin Identification of N-Glycosylated Proteins from the Central Nervous System of Drosophila Melanogaster Glycobiology, December 1, 2007; 17(12): 1388 - 1403. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Gutternigg, D. Kretschmer-Lubich, K. Paschinger, D. Rendic, J. Hader, P. Geier, R. Ranftl, V. Jantsch, G. Lochnit, and I. B. H. Wilson Biosynthesis of Truncated N-Linked Oligosaccharides Results from Non-orthologous Hexosaminidase-mediated Mechanisms in Nematodes, Plants, and Insects J. Biol. Chem., September 21, 2007; 282(38): 27825 - 27840. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Strasser, J. S. Bondili, J. Schoberer, B. Svoboda, E. Liebminger, J. Glossl, F. Altmann, H. Steinkellner, and L. Mach Enzymatic Properties and Subcellular Localization of Arabidopsis beta-N-Acetylhexosaminidases Plant Physiology, September 1, 2007; 145(1): 5 - 16. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Aoki, M. Perlman, J.-M. Lim, R. Cantu, L. Wells, and M. Tiemeyer Dynamic Developmental Elaboration of N-Linked Glycan Complexity in the Drosophila melanogaster Embryo J. Biol. Chem., March 23, 2007; 282(12): 9127 - 9142. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. J.A. Rouwendal, M. Wuhrer, D. E.A. Florack, C. A.M. Koeleman, A. M. Deelder, H. Bakker, G. M. Stoopen, I. van Die, J. P.F.G. Helsper, C. H. Hokke, et al. Efficient introduction of a bisecting GlcNAc residue in tobacco N-glycans by expression of the gene encoding human N-acetylglucosaminyltransferase III Glycobiology, March 1, 2007; 17(3): 334 - 344. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Cattaneo, M. E. Pasini, J. Intra, M. Matsumoto, F. Briani, M. Hoshi, and M. E. Perotti Identification and expression analysis of Drosophila melanogaster genes encoding {beta}-hexosaminidases of the sperm plasma membrane Glycobiology, September 1, 2006; 16(9): 786 - 800. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Tomiya, S. Narang, J. Park, B. Abdul-Rahman, O. Choi, S. Singh, J. Hiratake, K. Sakata, M. J. Betenbaugh, K. B. Palter, et al. Purification, Characterization, and Cloning of a Spodoptera frugiperda Sf9 beta-N-Acetylhexosaminidase That Hydrolyzes Terminal N-Acetylglucosamine on the N-Glycan Core J. Biol. Chem., July 14, 2006; 281(28): 19545 - 19560. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |