JBC Anatrace, Inc.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kornfeld, R.
Right arrow Articles by Canfield, W. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kornfeld, R.
Right arrow Articles by Canfield, W. M.

J Biol Chem, Vol. 273, Issue 36, 23203-23210, September 4, 1998

Purification and Multimeric Structure of Bovine N-Acetylglucosamine-1-phosphodiester alpha -N-Acetylglucosaminidase

Rosalind KornfeldDagger , Ming Baoparallel , Kevin Brewerparallel , Carolyn NollDagger , and William M. Canfieldparallel

From the Dagger  Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110 and the parallel  W. K. Warren Medical Research Institute and the Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104

N-Acetylglucosamine-1-phosphodiester alpha -N-Acetylglucosaminidase (EC 3.1.4.45; phosphodiester alpha -GlcNAcase) catalyzes the second step in the synthesis of the mannose 6-phosphate determinant required for efficient intracellular targeting of newly synthesized lysosomal hydrolases to the lysosome. A partially purified preparation of phosphodiester alpha -GlcNAcase from bovine pancreas was used to generate a panel of murine monoclonal antibodies. The anti-phosphodiester alpha -GlcNAcase monoclonal antibody UC1 was coupled to a solid support and used to immunopurify the bovine liver enzyme 670,000-fold in two steps to apparent homogeneity with an overall yield of 14%. The purified phosphodiester alpha -GlcNAcase has a specific activity of 498 µmol of [3H]GlcNAc-alpha -phosphomannose-alpha -methyl cleaved per h per mg of protein using 0.5 mM [3H]GlcNAc-alpha -phosphomannose-alpha -methyl as substrate.

The subunit structure of the enzyme was determined using a combination of analytical gel filtration chromatography, SDS-polyacrylamide gel electrophoresis, and amino-terminal sequencing. The data indicate that bovine phosphodiester alpha -GlcNAcase is a 272,000-Da complex of four identical 68,000-Da glycoprotein subunits arranged as two disulfide-linked homodimers. A soluble form of the enzyme, isolated from fetal bovine serum, showed the same subunit structure. Both forms of the enzyme reacted with a rabbit antibody raised to the amino-terminal peptide of the liver enzyme, suggesting that phosphodiester alpha -GlcNAcase is a type I membrane-spanning glycoprotein with its amino terminus in the lumen of the Golgi apparatus.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
J. Stockli, S. Honing, and J. Rohrer
The Acidic Cluster of the CK2 Site of the Cation-dependent Mannose 6-Phosphate Receptor (CD-MPR) but Not Its Phosphorylation Is Required for GGA1 and AP-1 Binding
J. Biol. Chem., May 28, 2004; 279(22): 23542 - 23549.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. Nair, B. E. Schaub, and J. Rohrer
Characterization of the Endosomal Sorting Signal of the Cation-dependent Mannose 6-Phosphate Receptor
J. Biol. Chem., June 27, 2003; 278(27): 24753 - 24758.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. Do, W.-S. Lee, P. Ghosh, T. Hollowell, W. Canfield, and S. Kornfeld
Human Mannose 6-Phosphate-uncovering Enzyme Is Synthesized as a Proenzyme That Is Activated by the Endoprotease Furin
J. Biol. Chem., August 9, 2002; 277(33): 29737 - 29744.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
W.-S. Lee, J. Rohrer, R. Kornfeld, and S. Kornfeld
Multiple Signals Regulate Trafficking of the Mannose 6-Phosphate-uncovering Enzyme
J. Biol. Chem., January 25, 2002; 277(5): 3544 - 3551.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Biol. CellHome page
J. Rohrer and R. Kornfeld
Lysosomal Hydrolase Mannose 6-Phosphate Uncovering Enzyme Resides in the trans-Golgi Network
Mol. Biol. Cell, June 1, 2001; 12(6): 1623 - 1631.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. Kornfeld, M. Bao, K. Brewer, C. Noll, and W. Canfield
Molecular Cloning and Functional Expression of Two Splice Forms of Human N-Acetylglucosamine-1-phosphodiester alpha -N-Acetylglucosaminidase
J. Biol. Chem., November 12, 1999; 274(46): 32778 - 32785.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. Ferlinz, G. Kopal, K. Bernardo, T. Linke, J. Bar, B. Breiden, U. Neumann, F. Lang, E. H. Schuchman, and K. Sandhoff
Human Acid Ceramidase. PROCESSING, GLYCOSYLATION, AND LYSOSOMAL TARGETING
J. Biol. Chem., September 14, 2001; 276(38): 35352 - 35360.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 1998 by the American Society for Biochemistry and Molecular Biology.