JBC Invitrogen Ultrasensitive Cytokine Assays

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 Hishiya, A.
Right arrow Articles by Nakamura, F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hishiya, A.
Right arrow Articles by Nakamura, F.

J Biol Chem, Vol. 274, Issue 38, 26705-26712, September 17, 1999

Protein Phosphatase 2C Inactivates F-actin Binding of Human Platelet Moesin

Akinori HishiyaDagger , Motoko Ohnishi, Shinri Tamura, and Fumihiko NakamuraDagger

From the Dagger  Department of Environmental Biology, Graduate School of Agricultural Science, Tohoku University, Sendai 981-8555, Japan and the  Department of Biochemistry, Institute of Development, Aging and Cancer, Tohoku University, Sendai 980-8575, Japan

During activation of platelets by thrombin phosphorylation of Thr558 in the C-terminal domain of the membrane-F-actin linking protein moesin increases transiently, and this correlates with protrusion of filopodial structures. Calyculin A enhances phosphorylation of moesin by inhibition of phosphatases. To measure this moesin-specific activity, a nonradioactive enzyme-linked immunosorbent assay method was developed with the synthetic peptide Cys-Lys555-Tyr-Lys-Thr(P)-Leu-Arg560 coupled to bovine serum albumin as the substrate and moesin phosphorylation state-specific polyclonal antibodies for the detection and quantitation of dephosphorylation. Calyculin A-sensitive and -insensitive protein-threonine phosphatase activities were detected in platelet lysates and separated by DEAE-cellulose chromatography. The calyculin A-sensitive enzyme was identified as a type 1 protein phosphatase. The calyculin A-insensitive enzyme activity was purified to homogeneity by phenyl- Sepharose, protamine-, and phosphonic acid peptide-agarose chromatography and characterized biochemically and immunologically as a 53-kDa protein(s) and a type 2C protein phosphatase (PP2C). Phosphorylation of Thr558 is necessary for F-actin binding of moesin in vitro. The purified enzyme, as well as bacterially made PP2Calpha and PP2Cbeta , efficiently dephosphorylate(s) highly purified platelet phospho-moesin. This reverses the activating effect of phosphorylation, and moesin no longer co-sediments with actin filaments. In vivo, regulation of these phosphatase activities are likely to influence dynamic interactions between the actin cytoskeleton and membrane constituents linked to moesin.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.


This article has been cited by other articles:


Home page
 Am. J. Physiol. Cell Physiol.Home page
L. Zhu, R. Zhou, S. Mettler, T. Wu, A. Abbas, J. Delaney, and J. G. Forte
High turnover of ezrin T567 phosphorylation: conformation, activity, and cellular function
Am J Physiol Cell Physiol, September 1, 2007; 293(3): C874 - C884.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
R. Zhou, L. Zhu, A. Kodani, P. Hauser, X. Yao, and J. G. Forte
Phosphorylation of ezrin on threonine 567 produces a change in secretory phenotype and repolarizes the gastric parietal cell
J. Cell Sci., October 1, 2005; 118(19): 4381 - 4391.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
R. Zhou, X. Cao, C. Watson, Y. Miao, Z. Guo, J. G. Forte, and X. Yao
Characterization of Protein Kinase A-mediated Phosphorylation of Ezrin in Gastric Parietal Cell Activation
J. Biol. Chem., September 12, 2003; 278(37): 35651 - 35659.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. Ofek, D. Ben-Meir, Z. Kariv-Inbal, M. Oren, and S. Lavi
Cell Cycle Regulation and p53 Activation by Protein Phosphatase 2Calpha
J. Biol. Chem., April 11, 2003; 278(16): 14299 - 14305.
[Abstract] [Full Text] [PDF]

Home page
 GeneticsHome page
A. J. Piekny, A. Wissmann, and P. E. Mains
Embryonic Morphogenesis in Caenorhabditis elegans Integrates the Activity of LET-502 Rho-Binding Kinase, MEL-11 Myosin Phosphatase, DAF-2 Insulin Receptor and FEM-2 PP2c Phosphatase
Genetics, December 1, 2000; 156(4): 1671 - 1689.
[Abstract] [Full Text]

Home page
 J. Biol. Chem.Home page
A. Cheng, P. Kaldis, and M. J. Solomon
Dephosphorylation of Human Cyclin-dependent Kinases by Protein Phosphatase Type 2Calpha and beta 2 Isoforms
J. Biol. Chem., October 27, 2000; 275(44): 34744 - 34749.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Biol.Home page
A. Gautreau, D. Louvard, and M. Arpin
Morphogenic Effects of Ezrin Require a Phosphorylation-induced Transition from Oligomers to Monomers at the Plasma Membrane
J. Cell Biol., July 11, 2000; 150(1): 193 - 204.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. A. Rosado, S. Jenner, and S. O. Sage
A Role for the Actin Cytoskeleton in the Initiation and Maintenance of Store-mediated Calcium Entry in Human Platelets. EVIDENCE FOR CONFORMATIONAL COUPLING
J. Biol. Chem., March 10, 2000; 275(11): 7527 - 7533.
[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 © 1999 by the American Society for Biochemistry and Molecular Biology.