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


     

Originally published In Press as doi:10.1074/mcp.T500024-MCP200 on December 11, 2005.
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental Data
Right arrow All Versions of this Article:
T500024-MCP200v1
5/4/749    most recent
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
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 arrowRequest Permissions
Right arrow Glossary
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kinoshita, E.
Right arrow Articles by Koike, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kinoshita, E.
Right arrow Articles by Koike, T.
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?

Molecular & Cellular Proteomics 5:749-757, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

Technology

Phosphate-binding Tag, a New Tool to Visualize Phosphorylated Proteins*,S

Eiji Kinoshita{ddagger},§,¶, Emiko Kinoshita-Kikuta{ddagger}, Kei Takiyama{ddagger} and Tohru Koike{ddagger},§,||

From the {ddagger} Department of Functional Molecular Science, Graduate School of Biomedical Sciences, and § Frontier Center for Microbiology, Hiroshima University, Kasumi 1-2-3, Hiroshima 734-8551, Japan

We introduce two methods for the visualization of phosphorylated proteins using alkoxide-bridged dinuclear metal (i.e. Zn2+ or Mn2+) complexes as novel phosphate-binding tag (Phos-tag) molecules. Both Zn2+- and Mn2+-Phos-tag molecules preferentially capture phosphomonoester dianions bound to Ser, Thr, and Tyr residues. One method is based on an ECL system using biotin-pendant Zn2+-Phos-tag and horseradish peroxidase-conjugated streptavidin. We demonstrate the electroblotting analyses of protein phosphorylation status by the phosphate-selective ECL signals. Another method is based on the mobility shift of phosphorylated proteins in SDS-PAGE with polyacrylamide-bound Mn2+-Phos-tag. Phosphorylated proteins in the gel are visualized as slower migration bands compared with corresponding dephosphorylated proteins. We demonstrate the kinase and phosphatase assays by phosphate affinity electrophoresis (Mn2+-Phos-tag SDS-PAGE).

To whom correspondence may be addressed. Tel.: 81-82-257-5281; Fax: 81-82-257-5336; E-mail: kinoeiji{at}hiroshima-u.ac.jp


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
 JCBHome page
T. Zimniak, K. Stengl, K. Mechtler, and S. Westermann
Phosphoregulation of the budding yeast EB1 homologue Bim1p by Aurora/Ipl1p
J. Cell Biol., August 10, 2009; 186(3): 379 - 391.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
A. H. Sikkema, S. H. Diks, W. F.A. den Dunnen, A. ter Elst, F. J.G. Scherpen, E. W. Hoving, R. Ruijtenbeek, P. J. Boender, R. de Wijn, W. A. Kamps, et al.
Kinome Profiling in Pediatric Brain Tumors as a New Approach for Target Discovery
Cancer Res., July 15, 2009; 69(14): 5987 - 5995.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
G. Bethke, T. Unthan, J. F. Uhrig, Y. Poschl, A. A. Gust, D. Scheel, and J. Lee
Flg22 regulates the release of an ethylene response factor substrate from MAP kinase 6 in Arabidopsis thaliana via ethylene signaling
PNAS, May 12, 2009; 106(19): 8067 - 8072.
[Abstract] [Full Text] [PDF]

Home page
 GENES CELLSHome page
Y. Hanyu, K. K. Imai, Y. Kawasaki, T. Nakamura, Y. Nakaseko, K. Nagao, A. Kokubu, M. Ebe, A. Fujisawa, T. Hayashi, et al.
Schizosaccharomyces pombe cell division cycle under limited glucose requires Ssp1 kinase, the putative CaMKK, and Sds23, a PP2A-related phosphatase inhibitor
Genes Cells, May 1, 2009; 14(5): 539 - 554.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
E. Ihara, P. L. Beck, M. Chappellaz, J. Wong, S. A. Medlicott, and J. A. MacDonald
Mitogen-Activated Protein Kinase Pathways Contribute to Hypercontractility and Increased Ca2+ Sensitization in Murine Experimental Colitis
Mol. Pharmacol., May 1, 2009; 75(5): 1031 - 1041.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
T. Kawashima, Y. C. Bao, Y. Minoshima, Y. Nomura, T. Hatori, T. Hori, T. Fukagawa, T. Fukada, N. Takahashi, T. Nosaka, et al.
A Rac GTPase-Activating Protein, MgcRacGAP, Is a Nuclear Localizing Signal-Containing Nuclear Chaperone in the Activation of STAT Transcription Factors
Mol. Cell. Biol., April 1, 2009; 29(7): 1796 - 1813.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
A. Grob, P. Roussel, J. E. Wright, B. McStay, D. Hernandez-Verdun, and V. Sirri
Involvement of SIRT7 in resumption of rDNA transcription at the exit from mitosis
J. Cell Sci., February 15, 2009; 122(4): 489 - 498.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
K. Takeya, K. Loutzenhiser, M. Shiraishi, R. Loutzenhiser, and M. P. Walsh
A highly sensitive technique to measure myosin regulatory light chain phosphorylation: the first quantification in renal arterioles
Am J Physiol Renal Physiol, June 1, 2008; 294(6): F1487 - F1492.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Biol. CellHome page
A. Devault, E. Gueydon, and E. Schwob
Interplay between S-Cyclin-dependent Kinase and Dbf4-dependent Kinase in Controlling DNA Replication through Phosphorylation of Yeast Mcm4 N-Terminal Domain
Mol. Biol. Cell, May 1, 2008; 19(5): 2267 - 2277.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. Tatematsu, N. Yoshimoto, T. Okajima, K. Tanizawa, and S. Kuroda
Identification of Ubiquitin Ligase Activity of RBCK1 and Its Inhibition by Splice Variant RBCK2 and Protein Kinase C{beta}
J. Biol. Chem., April 25, 2008; 283(17): 11575 - 11585.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
S.-i. Inoue, T. Kinoshita, M. Matsumoto, K. I. Nakayama, M. Doi, and K.-i. Shimazaki
Blue light-induced autophosphorylation of phototropin is a primary step for signaling
PNAS, April 8, 2008; 105(14): 5626 - 5631.
[Abstract] [Full Text] [PDF]

Home page
 DevelopmentHome page
H. Oh and K. D. Irvine
In vivo regulation of Yorkie phosphorylation and localization
Development, March 15, 2008; 135(6): 1081 - 1088.
[Abstract] [Full Text] [PDF]

Home page
 Plant Physiol.Home page
R. G. Uhrig, B. O'Leary, H. E. Spang, J. A. MacDonald, Y.-M. She, and W. C. Plaxton
Coimmunopurification of Phosphorylated Bacterial- and Plant-Type Phosphoenolpyruvate Carboxylases with the Plastidial Pyruvate Dehydrogenase Complex from Developing Castor Oil Seeds
Plant Physiology, March 1, 2008; 146(3): 1346 - 1357.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
E. Kinoshita-Kikuta, Y. Aoki, E. Kinoshita, and T. Koike
Label-free Kinase Profiling Using Phosphate Affinity Polyacrylamide Gel Electrophoresis
Mol. Cell. Proteomics, February 1, 2007; 6(2): 356 - 366.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Journal of Biological Chemistry 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2006 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement