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Major soluble proteins of grapevine ripe berries were extracted from six different cultivars including non vinifera, with trichloroacetic acid acetone and resolved in two‐dimensional electrophoresis (2‐DE) gels. About three hundred spots were detected on the 2‐DE map after colloidal blue staining. From 2‐DE map of cv. Gamay mesocarp, 67 proteins were identified (p > 0.95) using matrix‐assisted laser desorption/ionization‐mass spectrometry analysis. About 34%, 19%, and 13% of identified proteins play, respectively, a role in energy metabolism, defense, and stress response and primary metabolism. 2‐DE analysis revealed considerable accumulation of dehydrin, invertase, and a putative transcription factor in the ripe fruit, in addition to pathogenesis‐related proteins such as chitinase and thaumatin‐like proteins previously documented as prevalent proteins in ripe berries. Actual translation of redundant transcripts of unclear function such as Grip31, Grip32, and Grip61 recently cloned in ripe grape berries was confirmed. The relative abundance of UDP‐glucose pyrophosphorylase and vacuolar invertase strongly supported a key role of the apoplastic pathway of sugar loading during ripening. Comparative analysis shows that differences between cultivars were low, but different isoforms of alcohol dehydrogenase and of a transcription factor of hexose transporter were obvious in the six cultivars. Peptide mass fingerprinting suggests that the Adh isoforms would be Adh2/Adh6 or Adh2/Adh7 dimers and unambiguously shows that considerable deletion/insertion inside Adh7 are not cloning artifacts.