Glycobiology Advance Access originally published online on January 12, 2004
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Glycobiology vol 14 no 4 pp. 301-310, 2004
Glycobiology vol. 14 no. 4 © Oxford University Press 2004; all rights reserved.
Synthesis of a novel photoaffinity derivative of 1-deoxynojirimycin for active site-directed labeling of glucosidase I
Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742
Received on July 2, 2003; revised on November 13, 2003; accepted on November 14, 2004
Glucosidase I releases the distal 
1,2-glucosyl residue in the Glc3Man9GlcNAc2 precursor immediately after its transfer from the dolichol-P-P-linked intermediate in the endoplasmic reticulum and triggers the processes for the posttranslational remodeling, folding, and maturation of N-linked glycoproteins. The enzyme has been purified and characterized from several eukaryotic systems. Its cDNA and the gene have also been cloned. The enzyme is a target for the development of drugs for several pathological conditions. A structural analysis on the biochemically purified enzyme has been hampered because of its low abundance and unstable character. The recombinant enzyme has not been obtained in quantity and characterized. Glucosidase I is strongly inhibited by the glucose analog 1-deoxynojirimycin (DNM). To gain an insight into the architecture of the active site of the enzyme, we here report the synthesis of a photoactive derivative of DNM, viz. 4-(
-azidosalicylamido)butyl-5-amido-pentyl-1-DNM (ASBA-P-DNM). With an IC50 of 0.42 µM, it is nearly nine times stronger inhibitor than DNM (IC50 = 3.5 µM). On photolysis, the bound [125I]ASBA-P-DNM specifically labels the native enzyme, which yields a 24-kDa peptide after treatment with V8 protease, apparently representing the region around its active site. Thus ASBA-P-DNM should serve as a novel reagent to conduct structure–function analysis on glucosidase I.
1 To whom correspondence should be addressed; e-mail: ivijay{at}umd.edu
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  B. P Rempel and S. G Withers Covalent inhibitors of glycosidases and their applications in biochemistry and biology Glycobiology, August 1, 2008; 18(8): 570 - 586. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. S. P. Gowda, S. V. Madhunapantula, R. N. Achur, M. Valiyaveettil, V. P. Bhavanandan, and D. C. Gowda Structural Basis for the Adherence of Plasmodium falciparum-infected Erythrocytes to Chondroitin 4-Sulfate and Design of Novel Photoactivable Reagents for the Identification of Parasite Adhesive Proteins J. Biol. Chem., January 12, 2007; 282(2): 916 - 928. [Abstract] [Full Text] [PDF]
|
|