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The biochemical properties of the mitochondrial thiamine pyrophosphate carrier from Drosophila melanogaster
Author(s) -
Iacopetta Domenico,
Carrisi Chiara,
De Filippis Giuseppina,
Calcagnile Valeria M.,
Cappello Anna R.,
Chimento Adele,
Curcio Rosita,
Santoro Antonella,
Vozza Angelo,
Dolce Vincenza,
Palmieri Ferdinando,
Capobianco Loredana
Publication year - 2010
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2009.07550.x
Subject(s) - thiamine pyrophosphate , pyrophosphate , biochemistry , drosophila melanogaster , biology , mitochondrion , saccharomyces cerevisiae , melanogaster , thiamine , mitochondrial carrier , cofactor , yeast , enzyme , gene , bacterial outer membrane , escherichia coli
The mitochondrial carriers are a family of transport proteins that shuttle metabolites, nucleotides and cofactors across the inner mitochondrial membrane. The genome of Drosophila melanogaster encodes at least 46 members of this family. Only five of these have been characterized, whereas the transport functions of the remainder cannot be assessed with certainty. In the present study, we report the functional identification of two D. melanogaster genes distantly related to the human and yeast thiamine pyrophosphate carrier ( TPC ) genes as well as the corresponding expression pattern throughout development. Furthermore, the functional characterization of the D. melanogaster mitochondrial thiamine pyrophosphate carrier protein ( Dm Tpc1p) is described. Dm Tpc1p was over‐expressed in bacteria, the purified protein was reconstituted into liposomes, and its transport properties and kinetic parameters were characterized. Reconstituted Dm Tpc1p transports thiamine pyrophosphate and, to a lesser extent, pyrophosphate, ADP, ATP and other nucleotides. The expression of Dm Tpc1p in Saccharomyces cerevisiae TPC1 null mutant abolishes the growth defect on fermentable carbon sources. The main role of Dm Tpc1p is to import thiamine pyrophosphate into mitochondria by exchange with intramitochondrial ATP and/or ADP.