Glycobiology Advance Access published online on September 23, 2007
Glycobiology, doi:10.1093/glycob/cwm097
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Identification of N-glycosylated proteins from the central nervous system of Drosophila melanogaster
1 Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843-2128
2 Complex Carbohydrate Research Center, Department of Chemistry and Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602
3 Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia 30602
* To whom correspondence should be addressed: Tel. 979-458-4630, Fax: 979-845-9274, E-mail: panin{at}tamu.edu, Tel. 706-542-7806, Fax: 706-542-4412, E-mail: lwells{at}ccrc.uga.edu and Tel. 706-542-2740, Fax: 706-542-4412, E-mail: mtiemeyer{at}ccrc.uga.edu
Received on July 20, 2007; accepted on August 31, 2007
Although the function of many glycoproteins in the nervous system of fruit flies is well understood, information about the glycosylation profile and glycan attachment sites for such proteins is scarce. In order to fill this gap and to facilitate the analysis of N-linked glycosylation in the nervous system, we have performed an extensive survey of membrane-associated glycoproteins and their N-glycosylation sites isolated from the adult Drosophila brain. Following subcellular fractionation and trypsin digestion, we used different lectin affinity chromatography steps to isolate N-glycosylated glycopeptides. We identified a total of 205 glycoproteins carrying N-linked glycans and revealed their 307 N-glycan attachment sites. The size of the resulting dataset furthermore allowed the statistical characterization of amino acid distribution around the N-linked glycosylation sites. Glycan profiles were analyzed separately for glycopeptides that were strongly and weakly bound to Concanavalin A, or that failed to bind Concanavalin A but did bind to wheat germ agglutinin. High- or paucimannosidic glycans dominated each of the profiles, although the wheat germ agglutinin-bound glycan population was enriched in more extensively processed structures. A sialylated glycan structure was unambiguously detected in the wheat germ agglutinin-bound fraction. Despite the large amount of starting material, insufficient amount of glycopeptides was retained by the Wisteria floribunda and Sambucus nigra columns to allow glycan or glycoprotein identification, providing further evidence that the vast majority of glycoproteins in the adult Drosophila brain carry primarily high-mannose, paucimannose, and hybrid glycans. The obtained results should facilitate future genetic and molecular approaches addressing the role of N-glycosylation in the CNS of Drosophila.
CiteULike 
Connotea 
Del.icio.us 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]
|
|
|
|
 |
|
 C. L. R. Merry and C. M. Ward A sugar rush for developmental biology Development, April 15, 2008; 135(8): 1389 - 1393. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. S. Kang, J. Frank, C. H. Kang, H. Kajiura, M. Vikram, A. Ueda, S. Kim, J. D. Bahk, B. Triplett, K. Fujiyama, et al. Salt tolerance of Arabidopsis thaliana requires maturation of N-glycosylated proteins in the Golgi apparatus PNAS, April 15, 2008; 105(15): 5933 - 5938. [Abstract] [Full Text] [PDF]
|
|