Identification of phosphorylation and acetylation sites in αA-crystallin of the eye lens (mus musculus) after two-dimensional gel electrophoresis

Schaefer, Heike; Marcus, Katrin; Sickmann, Albert; Herrmann, Marion; Klose, Joachim; Meyer, Helmut E.
August 2003
Analytical & Bioanalytical Chemistry;Aug2003, Vol. 376 Issue 7, p966
Academic Journal
Posttranslational modifications are of great interest because of their relevance in biological systems as proteins are commonly activated or deactivated by phosphorylation, glycation and acetylation [1, 2]. During eye lens aging the number of the αA-crystallin isoproteins increases. This could be observed by the use of 2D-PAGE (two-dimensional gel electrophoresis). The number of αA-crystallin spots in the gel increased during eye lens aging. For further analysis the spots of 2D-PAGE were cut out and the identification of the proteins was done using nanoLC-ESI-MS/MS (liquid chromatography electrospray ionization tandem mass spectrometry). The created MS/MS-data were analyzed using the Sequest algorithm. Searches with different parameters were done to preferably get the complete sequence coverage and to identify posttranslational modifications of the αA-crystallin. The acetylated N-terminus of this protein could be detected. Furthermore, phosphorylation of serine 122 and 148 was identified in two different spots.


Related Articles

  • AutoMotif Server for prediction of phosphorylation sites in proteins using support vector machine: 2007 update. Dariusz Plewczynski; Adrian Tkacz; Leszek Rychlewski; Krzysztof Ginalski // Journal of Molecular Modeling;Jan2008, Vol. 14 Issue 1, p69 

    Abstract  We present here the recent update of AutoMotif Server (AMS 2.0) that predicts post-translational modification sites in protein sequences. The support vector machine (SVM) algorithm was trained on data gathered in 2007 from various sets of proteins containing experimentally...

  • Phosphorylation and ATP-binding induced conformational changes in the PrkC, Ser/Thr kinase from B. subtilis. Gruszczyński, Paweł; Obuchowski, Michał; Kaźmierkiewicz, Rajmund // Journal of Computer-Aided Molecular Design;Sep2010, Vol. 24 Issue 9, p733 

    Recent studies on the PrkC, serine-threonine kinase show that that the enzyme is located at the inner membrane of endospores and is responsible for triggering spore germination. The activity of the protein increases considerably after phosphorylation of four threonine residues placed on the...

  • The Helicobacter pylori CagA protein induces cortactin dephosphorylation and actin rearrangement by c-Src inactivation. Selbach, Matthias; Moese, Stefan; Hurwitz, Robert; Hauck, Christof R.; Meyer, Thomas F.; Backert, Steffen // EMBO Journal;2/1/2003, Vol. 22 Issue 3, p515 

    The gastric pathogen Helicobacter pylori translocates the CagA protein into epithelial cells by a type IV secretion process. Translocated CagA is tyrosine phosphorylated (CagAP-Tyr) on specific EPIYA sequence repeats by Src family tyrosine kinases. Phosphorylation of CagA induces the...

  • D4476, a cell-permeant inhibitor of CK1, suppresses the site-specific phosphorylation and nuclear exclusion of FOXO1a. Rena, Graham; Bain, Jenny; Elliott, Matthew; Cohen, Philip // EMBO Reports;Jan2004, Vol. 5 Issue 1, p60 

    The protein kinase CK1 phosphorylates serine residues that are located close to another phosphoserine in the consensus pSer-Xaa-Xaa-Ser. This specificity generates regions in its target proteins containing two or more neighbouring phosphoserine residues, termed here multisite phosphorylation...

  • Phosphorylation of the discs large tumour suppressor protein controls its membrane localisation and enhances its susceptibility to HPV E6-induced degradation. Massimi, P.; Narayan, N.; Cuenda, A.; Banks, L. // Oncogene;7/20/2006, Vol. 25 Issue 31, p4276 

    The Discs Large (Dlg) protein is intimately involved in regulating cell polarity and cell proliferation, and is targeted by the high-risk Human Papillomavirus (HPV) E6 proteins for proteasome-mediated degradation. We show here that exposure of cells to osmotic shock induces the...

  • MAPKAPK-2 AND SERINE PHOSPHORYLATION OF H5P25/27 AND αB-CRYSTALLIN DO NOT INCREASE MYOCARDIAL RESISTANCE TO INFARCTION. Gorog, D. A.; Tanno, M.; Fisher, S. G.; Cao, X. B.; Bellahcene, M.; Dighe, K.; Kabir, A. M. N.; Quinlan, R. A.; Kato, K.; Gaestel, M.; Marber, M. S. // Heart;May2004 Supplement 2, Vol. 90, pA24 

    This article focuses on a study related to MAPKAPK-2 and serine phosphorylation of HSP25/27 and αB-crystallin do not increase myocardial resistance to infarction. Ischaemic activation of MAPKAPK-2 and the phosphoryiation of its downstream targets H5P25/27 (pHSP25/27) and αB-crystallin may...

  • A rapid method for determining protein kinase phosphorylation specificity. Hutti, Jessica E.; Jarrell, Emily T.; Chang, James D.; Abbott, Derek W.; Storz, Peter; Toker, Alex; Cantley, Lewis C.; Turk, Benjamin E. // Nature Methods;Oct2004, Vol. 1 Issue 1, p27 

    Selection of target substrates by protein kinases is strongly influenced by the amino acid sequence surrounding the phosphoacceptor site. Identification of the preferred peptide phosphorylation motif for a given kinase permits the production of efficient peptide substrates and greatly simplifies...

  • Mass spectrometrical characterisation of mouse and rat synapsin isoforms 2a and 2b. Sung Ung Kang; Ming Zhang; Burgos, Miguel; Lubec, Gert // Amino Acids;Apr2010, Vol. 38 Issue 4, p1131 

    Synapsin 2 proteins are key elements of the synaptic machinery and still hold the centre stage in neuroscience research. Although fully sequenced at the nucleic acid level in mouse and rat, structural information on amino acid sequences and post-translational modifications (PTMs) is limited....

  • ITAM. Vohr, Hans-Werner // Encyclopedic Reference of Immunotoxicology;2005, p361 

    An entry for "immunoreceptor tyrosine-based activation motifs" is presented. It is composed of tyrosine residues separated by 9-12 amino acids. The phosphorylation of these residues leads to the activation of a number of effector functions.


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics