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

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Abstract.

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.

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Acknowledgements.

This work is part of the BMBF-project "Human Brain Proteome Project". Financial support from the BMBF (grant 031U102F) is gratefully acknowledged.

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Authors and Affiliations

  1. Medical Proteom-Center, Ruhr-University of Bochum, Bldg. ZKF E.141, Universitaetsstr. 150, 44801 , Bochum, Germany

    Heike Schaefer, Katrin Marcus, Albert Sickmann & Helmut E. Meyer

  2. Charité, Institute of Human Genetics, Humboldt-University, Campus Virchow-Klinikum, Augustenburger Platz 1, 13353, Berlin, Germany

    Marion Herrmann & Joachim Klose

Authors
  1. Heike Schaefer
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  2. Katrin Marcus
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  3. Albert Sickmann
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  4. Marion Herrmann
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  5. Joachim Klose
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  6. Helmut E. Meyer
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Correspondence to Helmut E. Meyer.

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Schaefer, H., Marcus, K., Sickmann, A. et al. Identification of phosphorylation and acetylation sites in αA-crystallin of the eye lens (mus musculus) after two-dimensional gel electrophoresis. Anal Bioanal Chem 376, 966–972 (2003). https://doi.org/10.1007/s00216-003-1983-1

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