Detection of protein phosphorylation on SDS-PAGE using probes with a phosphate-sensitive emission response

Bioconjugate Chemistry

Volumn 20, Issue 4, 2009, Pages 804-807

  Riechers, Alexander ^a   Schmidt, Florian ^a   Stadlbauer, Stefan ^a   Konig, Burkhard ^a  

^a UNIVERSITY OF REGENSBURG   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

ALPHA CASEIN; AMINO ACID; PHOSPHATE;

ARTICLE; BINDING SITE; ELECTRICITY; FIELD EMISSION; MOLECULAR PROBE; POLYACRYLAMIDE GEL ELECTROPHORESIS; PROTEIN BINDING; PROTEIN PHOSPHORYLATION; SENSITIVITY ANALYSIS;

EID: 65249086208     PISSN: 10431802     EISSN: None     Source Type: Journal    
DOI: 10.1021/bc9000307     Document Type: Article

References (22)

1. Pío, R., Martinez, A., Unsworth, E. J., Kowalak, J. A., Bengoechea, J. A., Zipfel, P. F., Elsasser, T. H., and Cuttitta, F. (2001) Complement factor H is a serum-binding protein for adrenomedullin, and the resulting complex modulates the bio-activities of both partners. J. Biol. Chem. 276, 12292-12300.
2. Misenheimer, T. M., Hahr, A. J., Harms, A. C., Annis, D. S., and Mosher, D. F. (2001) Disulfide connectivity of recombinant C-terminal region of human thrombospondin 2. J. Biol. Chem. 276, 45882-45887.
3. Williams, N. K., Prosselkov, P., Liepinsh, E., Line, I., Sharipo, A., Littler, D. R., Curmi, P. M. G., Otting, G., and Dixon, N. E. (2002) In vivo protein cyclization promoted by a circularly permuted Synechocystis sp. PCC6803 DnaB mini-intein. J. Biol. Chem. 277, 7790-7798.
4. Schulenberg, B., Goodman, T. N., Aggeler, R., Capaldi, R. A., and Patton, W. F. (2004) Characterization of dynamic and steady-state protein phosphorylation using a fluorescent phosphoprotein gel stain and mass spectrometry. Electrophoresis 24, 2526-2532.
5. Kinoshita, E., Kinoshita-Kikuta, E., Takiyama, K., and Koike, T. (2006) Phosphate-binding tag, a new tool to visualize phosphorylated proteins. Mol. Cell. Proteomics 5, 749-757.
6. Hunter, T. (1995) Protein kinases and phosphatases: The yin and yang of protein phosphorylation and signalling. Cell 80, 225-236.
7. Hunter, T. (2000) Signaling-2000 and beyond. Cell 100, 113-127.
8. Manning, G., Whyte, D. B., Martinez, R., Hunter, T., and Sudarsanam, S. (2002) The protein kinase complement of the human genome. Science 298 1912-1934.
9. Salomon, A. R., Ficarro, S. B., Brill, L. M., Brinker, A., Phung, Q. T., Ericson, C., Sauer, K., Brock, A., Horn, D. M., Schultz, P. G., and Peters, E. C. (2003) Profiling of tyrosine phosphorylation pathways in human cells using mass spectrometry. Proc. Natl. Acad. Sci. U.S.A. 100, 443-448.
10. Conrads, T. P., and Veenstra, T. D. (2005) An enriched look at tyrosine phosphorylation. Nat. Biotechnol. 23, 36-37.
11. Larsen, M. R., Sorensen, G. L., Fey, S. J., Larsen, P. M., and Roepstorff, P. (2001) Phospho-proteomics: Evaluation of the use of enzymatic de-phosphorylation and differential mass spectrometric peptide mass mapping for site specific phosphorylation assignment in proteins separated by gel electrophoresis. Proteomics 1, 223-238.
12. Immler, D., Gremm, D., Kirsch, D., Spengler, B., Presek, P., and Meyer, H. E. (1998) Identification of phosphorylated proteins from thrombin-activated human platelets isolated by two-dimensional gel electrophoresis by electrospray ionization-tandem mass spectrometry (ESI-MS/MS) and liquid chromatography- electrospray ionization-mass spectrometry (LC-ESI-MS). Elec-trophoresis 19, 1015-1023.
13. Ojida, A., Kohira, T., and Hamachi, I. (2004) Phosphoprotein-selective recognition and staining in SDS-PAGE by bis-Zn(II)-dipycolylamine-appended anthracene. Chem. Lett. 33, 1024-1025.
14. Ojida, A., Mito-oka, Y., Inoue, M., and Hamachi, I. (2002) First artificial receptors and chemosensors toward phosphorylated peptide in aqueous solution. J. Am. Chem. Soc. 124, 6256-6258.
15. Grauer, A., Riechers, A., Ritter, S., and Konig, B. (2008) Synthetic receptors for the differentiation of phosphorylated peptides with nanomolar affinities. Chem. Eur. J. 14, 8922-8927.
16. Mizukami, S., Nagano, T., Urano, N., Odani, Y., and Kikuchi, K. (2002) A fluorescent anion sensor that works in neutral aqueous solution for bioanalytical application. J. Am. Chem. Soc. 124, 3920-3925.
17. Wolfbeis, O. S., and Uray, G. (1978) Fluoreszenzspektren, Photodissoziation und Phototautomerie einiger 4-Hydroxycumarine. Monatsh. Chem. 109, 123-136.
18. To assess the influence of the fluorophore further, we also prepared the same type of probe with pyrene and dansyl fluorophores. Neither of these probes exhibited any distinction between phosphorylated and nonphosphorylated proteins (data not shown).
19. Kruppa, M., and Konig, B. (2006) Reversible coordinative bonds in molecular recognition. Chem. Rev. 106, 3520-3560.
20. Kruppa, M., Frank, D., Leffler-Schuster, H., and Konig, B. (2006) Screening of metal complex-amino acid side chain interactions by potentiometric titration. Inore. Chim. Acta 359, 1159-1168.
21. Lugina, L. N., Davidenko, N. K., and Gavrish, S. P. (1985) Quenching of the fluorescence of riboflavin by the histidinates and alaninates of nickel, copper, and zinc. Theor. Exp. Chem. 21, 466-170.
22. Hou, J. L., Yi, H. P., Shao, X.-B., Li, C., Wu, Z.-Q., Jiang, X.-K., Wu, L.-Z., Tung, C.-H., and Li, Z.-T. (2004) In situ click chemistry: enzyme-generated inhibitors of carbonic anhydrase II. Angew. Chem. 117, 118-122.