|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 275, Issue 38, 29488-29502, September 22, 2000
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From the Department of Biochemistry and Molecular Genetics, Israel
Institute for Biological Research, Ness-Ziona 74100, Israel
The tetrameric form of native serum-derived
bovine acetylcholinesterase is retained in the circulation for much
longer periods (mean residence time, MRT = 1390 min) than
recombinant bovine acetylcholinesterase (rBoAChE) produced in the
HEK-293 cell system (MRT = 57 min). Extensive matrix-assisted
laser desorption ionization-time of flight analyses established that
the basic structures of the N-glycans associated with the
native and recombinant enzymes are similar (the major species
(50-60%) are of the biantennary fucosylated type and 20-30% are of
the triantennary type), yet the glycan termini of the native enzyme are
mostly capped with sialic acid (82%) and
Hierarchy of Post-translational Modifications Involved in the
Circulatory Longevity of Glycoproteins
DEMONSTRATION OF CONCERTED CONTRIBUTIONS OF GLYCAN SIALYLATION
AND SUBUNIT ASSEMBLY TO THE PHARMACOKINETIC BEHAVIOR OF BOVINE
ACETYLCHOLINESTERASE*
-galactose (12%), whereas
glycans of the recombinant enzyme exhibit a high level of exposed
-galactose residues (50%) and a lack of -galactose. Glycan
termini of both fetal bovine serum and rBoAChE were altered in
vitro using exoglycosidases and sialyltransferase or in
vivo by a HEK-293 cell line developed specifically to allow
efficient sialic acid capping of -galactose-exposed termini. In
addition, the dimeric and monomeric forms of rBoAChE were
quantitatively converted to tetramers by complexation with a synthetic
peptide representing the human ColQ-derived proline-rich attachment
domain. Thus by controlling both the level and nature of
N-glycan capping and subunit assembly, we generated and
characterized 9 distinct bovine AChE glycoforms displaying a 400-fold
difference in their circulatory lifetimes (MRT = 3.5-1390 min).
This revealed some general rules and a hierarchy of post-translation
factors determining the circulatory profile of glycoproteins.
Accordingly, an rBoAChE was generated that displayed a circulatory
profile indistinguishable from the native form.
*
This work was supported by the United States Army Research
and Development Command, Contracts DAMD17-96-C-6088 and
DAMD17-00-C-0021 (to A. S.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 972-8-9381595;
Fax: 972-8-9401404; E-mail: avigdor@iibr.gov.il.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  O. Cohen, C. Kronman, A. Lazar, B. Velan, and A. Shafferman Controlled Concealment of Exposed Clearance and Immunogenic Domains by Site-specific Polyethylene Glycol Attachment to Acetylcholinesterase Hypolysine Mutants J. Biol. Chem., December 7, 2007; 282(49): 35491 - 35501. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Evron, B. C. Geyer, I. Cherni, M. Muralidharan, J. Kilbourne, S. P. Fletcher, H. Soreq, and T. S. Mor Plant-derived human acetylcholinesterase-R provides protection from lethal organophosphate poisoning and its chronic aftermath FASEB J, September 1, 2007; 21(11): 2961 - 2969. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Cohen, C. Kronman, L. Raveh, O. Mazor, A. Ordentlich, and A. Shafferman Comparison of Polyethylene Glycol-Conjugated Recombinant Human Acetylcholinesterase and Serum Human Butyrylcholinesterase as Bioscavengers of Organophosphate Compounds Mol. Pharmacol., September 1, 2006; 70(3): 1121 - 1131. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Sankala, A. Brannstrom, T. Schulthess, U. Bergmann, E. Morgunova, J. Engel, K. Tryggvason, and T. Pikkarainen Characterization of Recombinant Soluble Macrophage Scavenger Receptor MARCO J. Biol. Chem., August 30, 2002; 277(36): 33378 - 33385. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. G. Duysen, C. F. Bartels, and O. Lockridge Wild-Type and A328W Mutant Human Butyrylcholinesterase Tetramers Expressed in Chinese Hamster Ovary Cells Have a 16-Hour Half-Life in the Circulation and Protect Mice from Cocaine Toxicity J. Pharmacol. Exp. Ther., August 1, 2002; 302(2): 751 - 758. [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 |