JBC Ideal method for primary cell transfection

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 An addition or correction has been published
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 Namchuk, M.
Right arrow Articles by Baekkeskov, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Namchuk, M.
Right arrow Articles by Baekkeskov, S.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Volume 272, Number 3, Issue of January 17, 1997 pp. 1548-1557
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Phosphorylation of Serine Residues 3, 6, 10, and 13 Distinguishes Membrane Anchored from Soluble Glutamic Acid Decarboxylase 65 and Is Restricted to Glutamic Acid Decarboxylase 65alpha

(Received for publication, August 1, 1996, and in revised form, October 25, 1996)

Mark Namchuk Dagger , LeAnn Lindsay Dagger , Christoph W. Turck , Jamil Kanaani Dagger and Steinunn Baekkeskov Dagger

From the Dagger  Department of Medicine, Department of Microbiology and Immunology, and Hormone Research Institute and the  Howard Hughes Medical Institute, and Department of Medicine, University of California San Francisco, San Francisco, California 94143

GAD65, the smaller isoform of the gamma -aminobutyric acid-synthesizing enzyme glutamic acid decarboxylase is detected as an alpha /beta doublet of distinct mobility on SDS-polyacrylamide gel electrophoresis. Glutamic acid decarboxylase (GAD) 65 is reversibly anchored to the membrane of synaptic vesicles in neurons and synaptic-like microvesicles in pancreatic beta -cells. Here we demonstrate that GAD65alpha but not beta  is phosphorylated in vivo and in vitro in several cell types. Phosphorylation is not the cause of the alpha /beta heterogeneity but represents a unique post-translational modification of GAD65alpha . Two-dimensional protein analyses identified five phosphorylated species of three different charges, which are likely to represent mono-, di-, and triphosphorylated GAD65alpha in different combinations of phosphorylated serines. Phosphorylation of GAD65alpha was located at serine residues 3, 6, 10, and 13, shown to be mediated by a membrane bound kinase, and distinguish the membrane anchored, and soluble forms of the enzyme. Phosphorylation status does not affect membrane anchoring of GAD65, nor its Km or Vmax for glutamate. The results are consistent with a model in which GAD65alpha and -beta constitute the two subunits of the native GAD65 dimer, only one of which, alpha , undergoes phosphorylation following membrane anchoring, perhaps to regulate specific aspects of GAD65 function in the synaptic vesicle membrane.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Cell Sci.Home page
J. Kanaani, G. Patterson, F. Schaufele, J. Lippincott-Schwartz, and S. Baekkeskov
A palmitoylation cycle dynamically regulates partitioning of the GABA-synthesizing enzyme GAD65 between ER-Golgi and post-Golgi membranes
J. Cell Sci., February 15, 2008; 121(4): 437 - 449.
[Abstract] [Full Text] [PDF]

Home page
 J BiochemHome page
S. Matsukawa and H. Ueno
Expression of Glutamate Decarboxylase Isoform, GAD65, in Human Mononuclear Leucocytes: A Possible Implication of C-terminal End Deletion by Western Blot and RT PCR Study
J. Biochem., November 1, 2007; 142(5): 633 - 638.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
J. Kanaani, M. J. Diacovo, A. E.-D. El-Husseini, D. S. Bredt, and S. Baekkeskov
Palmitoylation controls trafficking of GAD65 from Golgi membranes to axon-specific endosomes and a Rab5a-dependent pathway to presynaptic clusters
J. Cell Sci., April 15, 2004; 117(10): 2001 - 2013.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. Fan, C. W. Turck, and R. Derynck
Characterization of Growth Factor-induced Serine Phosphorylation of Tumor Necrosis Factor-alpha Converting Enzyme and of an Alternatively Translated Polypeptide
J. Biol. Chem., May 9, 2003; 278(20): 18617 - 18627.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
H. Jin, H. Wu, G. Osterhaus, J. Wei, K. Davis, D. Sha, E. Floor, C.-C. Hsu, R. D. Kopke, and J.-Y. Wu
Demonstration of functional coupling between gamma -aminobutyric acid (GABA) synthesis and vesicular GABA transport into synaptic vesicles
PNAS, April 1, 2003; 100(7): 4293 - 4298.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Biol.Home page
J. Kanaani, A. E.-D. El-Husseini, A. Aguilera-Moreno, J. M. Diacovo, D. S. Bredt, and S. Baekkeskov
A combination of three distinct trafficking signals mediates axonal targeting and presynaptic clustering of GAD65
J. Cell Biol., September 29, 2002; 158(7): 1229 - 1238.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Kanaani, D. Lissin, S. F. Kash, and S. Baekkeskov
The Hydrophilic Isoform of Glutamate Decarboxylase, GAD67, Is Targeted to Membranes and Nerve Terminals Independent of Dimerization with the Hydrophobic Membrane-anchored Isoform, GAD65
J. Biol. Chem., December 24, 1999; 274(52): 37200 - 37209.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
N. Tian, C. Petersen, S. Kash, S. Baekkeskov, D. Copenhagen, and R. Nicoll
The role of the synthetic enzyme GAD65 in the control of neuronal gamma -aminobutyric acid release
PNAS, October 26, 1999; 96(22): 12911 - 12916.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C.-C. Hsu, C. Thomas, W. Chen, K. M. Davis, T. Foos, J. L. Chen, E. Wu, E. Floor, J. V. Schloss, and J.-Y. Wu
Role of Synaptic Vesicle Proton Gradient and Protein Phosphorylation on ATP-mediated Activation of Membrane-associated Brain Glutamate Decarboxylase
J. Biol. Chem., August 20, 1999; 274(34): 24366 - 24371.
[Abstract] [Full Text] [PDF]

Home page
 J. Neurosci.Home page
Y. Jin, E. Jorgensen, E. Hartwieg, and H. R. Horvitz
The Caenorhabditis elegans Gene unc-25 Encodes Glutamic Acid Decarboxylase and Is Required for Synaptic Transmission But Not Synaptic Development
J. Neurosci., January 15, 1999; 19(2): 539 - 548.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S. F. Kash, R. S. Johnson, L. H. Tecott, J. L. Noebels, R. D. Mayfield, D. Hanahan, and S. Baekkeskov
Epilepsy in mice deficient in the 65-kDa isoform of glutamic acid decarboxylase
PNAS, December 9, 1997; 94(25): 14060 - 14065.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C.-C. Hsu, K. M. Davis, H. Jin, T. Foos, E. Floor, W. Chen, J. B. Tyburski, C.-Y. Yang, J. V. Schloss, and J.-Y. Wu
Association of L-Glutamic Acid Decarboxylase to the 70-kDa Heat Shock Protein as a Potential Anchoring Mechanism to Synaptic Vesicles
J. Biol. Chem., June 30, 2000; 275(27): 20822 - 20828.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
B. Rubi, H. Ishihara, F. G. Hegardt, C. B. Wollheim, and P. Maechler
GAD65-mediated Glutamate Decarboxylation Reduces Glucose-stimulated Insulin Secretion in Pancreatic Beta Cells
J. Biol. Chem., September 21, 2001; 276(39): 36391 - 36396.
[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 © 1997 by the American Society for Biochemistry and Molecular Biology.