Originally published In Press as doi:10.1074/mcp.M700170-MCP200 on December 3, 2007.
Molecular & Cellular Proteomics 7:684-696, 2008.
© 2008 by The American Society for Biochemistry and Molecular Biology, Inc.
Special Issue: 8th International Symposium On Mass Spectrometry In The Life Sciences
Quantitative Analysis of Synaptic Phosphorylation and Protein Expression*,S
Jonathan C. Trinidad ,
Agnes Thalhammer ,
Christian G. Specht ,¶,
Aenoch J. Lynn ,
Peter R. Baker ,
Ralf Schoepfer ,|| and
Alma L. Burlingame ,**
From the Mass Spectrometry Facility, Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143 and Laboratory for Molecular Pharmacology, Department of Pharmacology, University College London, Gower Street, London WC1E 6BT, United Kingdom
The postsynaptic density (PSD) signaling machinery contains proteins with diverse functions. Brain region-specific variations in PSD components mediate distinct physiological responses to synaptic activation. We have developed mass spectrometry-based methods to comprehensively compare both relative protein expression and phosphorylation status from proteins present in biochemical preparations of postsynaptic density. Using these methods, we determined the relative expression of 2159 proteins and 1564 phosphorylation sites in PSD preparations from murine cortex, midbrain, cerebellum, and hippocampus. These experiments were conducted twice using independent biological replicates, which allowed us to assess the experimental and biological variability in this system. Concerning protein expression, cluster analysis revealed that known functionally associated proteins display coordinated synaptic expression. Therefore, proteins identified as co-clustering with known protein complexes are prime candidates for assignment as previously unrecognized components. Concerning degree of phosphorylation, we observed more extensive phosphorylation sites on N-methyl-D-aspartate (NMDA) receptors than -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors, consistent with the central role of N-methyl-D-aspartate receptors in processing synaptic transmission patterns. Average kinase and phosphatase levels were highest in the hippocampus, correlating with a higher overall phosphopeptide abundance present in this brain region. These findings suggest that the hippocampus utilizes reversible protein phosphorylation to a greater extent than other brain regions when modifying synaptic strength.
|| To whom correspondence may be addressed. E-mail: r.schoepfer{at}ucl.ac.uk
** To whom correspondence may be addressed. E-mail: alb{at}cgl.ucsf.edu

CiteULike Complore Connotea Del.icio.us Digg Reddit Technorati What's this?
This article has been cited by other articles:

|
 |

|
 |
 
R. J. Chalkley, A. Thalhammer, R. Schoepfer, and A. L. Burlingame
Identification of protein O-GlcNAcylation sites using electron transfer dissociation mass spectrometry on native peptides
PNAS,
June 2, 2009;
106(22):
8894 - 8899.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
D. Edbauer, D. Cheng, M. N. Batterton, C.-F. Wang, D. M. Duong, M. B. Yaffe, J. Peng, and M. Sheng
Identification and Characterization of Neuronal Mitogen-activated Protein Kinase Substrates Using a Specific Phosphomotif Antibody
Mol. Cell. Proteomics,
April 1, 2009;
8(4):
681 - 695.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
C. Wozny, J. Breustedt, F. Wolk, F. Varoqueaux, S. Boretius, A. R. Zivkovic, A. Neeb, J. Frahm, D. Schmitz, N. Brose, et al.
The function of glutamatergic synapses is not perturbed by severe knockdown of 4.1N and 4.1G expression
J. Cell Sci.,
March 1, 2009;
122(5):
735 - 744.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. Goudreault, L. M. D'Ambrosio, M. J. Kean, M. J. Mullin, B. G. Larsen, A. Sanchez, S. Chaudhry, G. I. Chen, F. Sicheri, A. I. Nesvizhskii, et al.
A PP2A Phosphatase High Density Interaction Network Identifies a Novel Striatin-interacting Phosphatase and Kinase Complex Linked to the Cerebral Cavernous Malformation 3 (CCM3) Protein
Mol. Cell. Proteomics,
January 1, 2009;
8(1):
157 - 171.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. O. Collins, L. Yu, I. Campuzano, S. G. N. Grant, and J. S. Choudhary
Phosphoproteomic Analysis of the Mouse Brain Cytosol Reveals a Predominance of Protein Phosphorylation in Regions of Intrinsic Sequence Disorder
Mol. Cell. Proteomics,
July 1, 2008;
7(7):
1331 - 1348.
[Abstract]
[Full Text]
[PDF]
|
 |
|
Copyright © 2008 by the American Society for Biochemistry and Molecular Biology.
|
Advertisement
Advertisement
|