Abstract
We have performed the iTRAQ mass spectrometry analysis of extracellular proteasomes and revealed posttranslational modifications (PTMs) of subunits of extracellular 26S proteasomes. There are novel sites of ubiquitination and acetylation found in the proteasome subunits α2 (K196), α4 (K189 and K234), α6 (K217), and Rpn6 (A2). We have revealed that the extracellular proteasomes contain the standard 26S proteasome subunits as well as the PA200 regulator, previously believed to be located exclusively in the cell nuclei. We have additionally performed a primary screening of proteins interacting with extracellular proteasomes and found that associated with proteasomes are the proteins participating in such major cellular processes as transcription, DNA repair and translation, as well as cytoskeleton proteins and the ubiquitinproteasome system (UPS). Using the method of co-purification and immunoblotting we have confirmed interaction of proteasomes with nine proteins randomly chosen from the list of the identified extracellular proteasome partners.
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Abbreviations
- SDS:
-
sodium dodecyl sulfate
- PAAG:
-
polyacrylamide gel
- PTM:
-
posttranslational modifications
- UPS:
-
ubiquitin-proteasome system
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Original Russian Text © Yu.Ya. Zaikova, V.A. Kulichkova, Yu.B. Ermolaeva, A. Bottrill, N.A. Barlev, A.S. Tsimokha, 2013, published in Tsitologiya, Vol. 55, No. 2, 2013, pp. 111–122.
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Zaikova, Y.Y., Kulichkova, V.A., Ermolaeva, Y.B. et al. Characterization of extracellular proteasomes and its interacting proteins by iTRAQ mass spectrometry. Cell Tiss. Biol. 7, 253–265 (2013). https://doi.org/10.1134/S1990519X13030139
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DOI: https://doi.org/10.1134/S1990519X13030139