The endoplasmic reticulum (ER) and the Golgi apparatus are crucial for the secretory pathway and play a central role in fruit ripening. The Golgi complex functions to posttranslationally modify newly synthesized proteins and sort them, packaged in vesicles for transport to their sites of function. We aim to monitor the pattern of protein expression along the secretory pathway during tomato fruit ripening. ER and Golgi microsomal vesicles were separated by centrifugation through different types of density gradients. Assuming that an organelle has a unique distribution pattern it is possible to identify ER or Golgi components by comparing their enrichment using known markers of these compartments. Profiling of protein distribution was determined by DIGE analysis of gradient fractions, respectively enriched in the ER and Golgi microsomes, to allow quantitative evaluation of differentially expressed proteins. Moreover, to overcome technical limitations resulting in reduced identifications of transmembrane proteins and posttranslational modifications, we developed an alternative multidimensional identification strategy using a combination of 1-D electrophoresis and nanoLC-ESIMS/MS.

ER AND GOLGI PROTEOME DYNAMICS DURING TOMATO FRUIT RIPENING

Maria Benedetta Mattei;
2010-01-01

Abstract

The endoplasmic reticulum (ER) and the Golgi apparatus are crucial for the secretory pathway and play a central role in fruit ripening. The Golgi complex functions to posttranslationally modify newly synthesized proteins and sort them, packaged in vesicles for transport to their sites of function. We aim to monitor the pattern of protein expression along the secretory pathway during tomato fruit ripening. ER and Golgi microsomal vesicles were separated by centrifugation through different types of density gradients. Assuming that an organelle has a unique distribution pattern it is possible to identify ER or Golgi components by comparing their enrichment using known markers of these compartments. Profiling of protein distribution was determined by DIGE analysis of gradient fractions, respectively enriched in the ER and Golgi microsomes, to allow quantitative evaluation of differentially expressed proteins. Moreover, to overcome technical limitations resulting in reduced identifications of transmembrane proteins and posttranslational modifications, we developed an alternative multidimensional identification strategy using a combination of 1-D electrophoresis and nanoLC-ESIMS/MS.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11697/126435
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