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Journal of Biochemistry Advance Access originally published online on May 31, 2008
Journal of Biochemistry 2008 144(3):389-392; doi:10.1093/jb/mvn076
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© 2008 The Japanese Biochemical Society

Identification of the Drosophila melanogaster Mitochondrial Citrate Carrier: Bacterial Expression, Reconstitution, Functional Characterization and Developmental Distribution

Chiara Carrisi1,{dagger}, Marianna Madeo1,2,{dagger}, Patrizia Morciano3, Vincenza Dolce2, Giovanni Cenci3, Anna Rita Cappello2, Giancarlo Mazzeo2, Domenico Iacopetta2 and Loredana Capobianco1,*

1Department of Biological and Environmental Sciences and Technologies, University of Salento, Lecce, Italy; 2Department of Pharmaco-Biology, University of Calabria, Arcavacata di Rende, Cosenza, Italy; and 3Department of Basic and Applied Biology, University of L’Aquila, L’Aquila, Italy

*To whom correspondence should be addressed. Tel: +39(0)832298864, Fax: +39(0)832298626, E-mail: loredana.capobianco{at}unile.it

Received May 9, 2008; Accepted May 27, 2008


  Abstract

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 four of them have been characterized: the two isoforms of the ADP/ATP translocase, the brain uncoupling protein and the carnitine/acylcarnitine carriers. The transport functions of the remainders cannot be assessed with certainty. One of them, the product of the gene CG6782, shows a fairly close sequence homology to the known sequence of the rat mitochondrial citrate carrier. In this article the fruit fly protein coding by the CG6782 gene has been functionally characterized by over-expression in Escherichia coli and reconstitution into liposomes. It shows to have similar transport properties of the eukaryotic mitochondrial citrate carriers previously biochemically characterized. This indicates that in addition to the protein sequence conservation, insect and mammalian citrate carriers are also significantly related at the functional level suggesting that Drosophila may be used as model organism for the study of mitochondrial solute transporter. The DmCIC expression pattern throughout development was also investigated; the transcripts were detected at equal levels in all stages analysed.

Key Words: citrate carrier, Drosophila melanogaster, metabolite transporter, mitochondria, transport

Abbreviations: BAT, Bathophenanthroline; BTA, benzenetricarboxylic acid; DmCIC, Drosophila melanogaster citrate carrier; PIPES, 1,4-piperazinediethanesulfonic acid; PLP, Pyridoxal 5'-phosphate; RT-PCR, reverse transcription polymerase chain reaction

{dagger}These two authors contributed equally to this work.


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