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RESEARCH PROGRAM
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Research Units
Similar research programs:
- 1 - Structure, function and biogenesis of mitochondrial transporters for metabolites
- 2 - Reconstitution into liposomes of plasma membrane amino acid transporters specific for glutamine: an experimental model for the study of the function and the regulation of native and over-expressed proeins.
- 3 - Structural and functional characterization of the human type 2 excitatory aminoacid transporter (EAAT2) and of its interaction with new ligands of pharmaceutical interest.
- 4 - Supramolecular complexes of sorcin in the generation and regulation of Calcium-dependent cellular functions
- 5 - Structural properties and functional activities in a chromatin remodeling nuclear protein complex
- 6 - Molecular analysis of erythropoiesis: Post-genomic and functional approach
- 7 - Identification of new mitochondrial carriers in man
- 8 - From the study of global regulation of gene expression to the study of virulence in Mycobacterium tuberculosis
- 9 - Identification of folding and misfolding determinants by site-directed mutagenesis.
- 10 - Physiology and pathology of protein folding, trafficking and signaling in the endoplasmic reticulum
Scientific and education field classification
- Field: Scienze biologiche
International Patent Classification
- CHEMISTRY; METALLURGY
- BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- ORGANIC CHEMISTRY (such compounds as the oxides, sulfides, or oxysulfides of carbon, cyanogen, phosgene, hydrocyanic acid or salts thereof C01; products obtained from layered base-exchange silicates by ion-exchange with organic compounds such as ammonium, phosphonium or sulfonium compounds or by intercalation of organic compounds C01B33/44; macromolecular compounds C08; dyes C09; fermentation products C12; fermentation or enzyme-using processes to synthesise a desired chemical compound or composition or to separate optical isomers from a racemic mixture C12P; production of organic compounds by electrolysis or electrophoresis C25B3/00, C25B7/00)
- PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Geographical classification
- Region: Lazio
Bibliografia
1. F. Palmieri (2004) Pflugers Arch. - Eur. J. Physiol. 447, 689-7092. E. Pebay-Peyroula, C. Dahout-Gonzalez , R. Kahn, V. Trezeguet, G.J. Lauquin, G. Brandolin (2003) Nature 426, 39-44
3. Z. Xie, E. Turk and E. M. Wright (2000) J. Biol. Chem. 275, 25959-25964
4. M. Sahin-Toth, A. Karlin and H.R. Kaback (2000) Proc. Natl. Acad. Sci. USA 97, 10729-10732
5. N. Tamura, S. Konishi, S. Iwaki, T. Kimura-Someya, S. Nada and A. Yamaguchi (2001) J. Biol. Chem. 276, 20330-20339
6. F. Palmieri, E. Quagliariello and M. Klingenberg (1972) Eur. J. Biochem. 29, 408-416.
7. R. Krämer and F. Palmieri in "Molecular Mechanisms in Bioenergetics" (L. Ernster ed.), Elsevier Science Publishers B.V., Amsterdam, (1992) 359-384
8. F. Bisaccia, C. Indiveri and F. Palmieri (1985) Biochim. Biophys. Acta 810, 362-369
9. C. Indiveri, F. Palmieri, F. Bisaccia and R. Krämer (1987) Biochim. Biophys. Acta, 890, 310-318
10. C. Indiveri, T. Dierks, R. Krämer and F. Palmieri (1991) Eur. J. Biochem. 198, 339-347
11. M.J. Runswick, J.E. Walker, F. Bisaccia, V. Iacobazzi and F. Palmieri (1990) Biochemistry 29, 11033-11040
12. V. Iacobazzi, F. Palmieri, M.J. Runswick and J.E. Walker (1992) DNA Sequence 3, 79-88
13. V. Dolce, A. Messina, A. Cambria and F. Palmieri (1994) DNA Sequence 5, 103-109
14. S. Piccininni, V. Iacobazzi, G. Lauria, M. Rocchi and F. Palmieri (1998) Cytogenet. Cell Genet. 83, 256-257
15. F. Palmieri (1994) FEBS Lett. 346, 48-54
16. F. Bisaccia, L. Capobianco, G. Brandolin and F. Palmieri (1994) Biochemistry 33, 3705-3713
17. G. Fiermonte, J.E. Walker and F. Palmieri (1993) Biochem. J. 294, 293-299
18. H. Wohlrab and C. Briggs (1994) Biochemistry 33, 9371-9375
19. R.S. Kaplan, J.A. Mayor, D.A. Gremse and D.O. Wood (1995) J. Biol. Chem. 270, 4108-4114
20. Y. Xu, J.A. Mayor, D. Gremse, D.O. Wood and R.S. Kaplan (1995) Biochem. Biophys. Res. Commun. 207, 783-789
21. G. Fiermonte, V. Dolce and F. Palmieri (1998) J. Biol. Chem. 273, 22782-22787
22. L. Palmieri, A. Vozza, G. Agrimi, V. De Marco, M.J. Runswick, F. Palmieri and J.E. Walker (1999) J. Biol. Chem. 274, 22184-22190
23. G. Fiermonte, V. Dolce, L. Palmieri, M. Ventura, M.J. Runswick, F. Palmieri and J.E. Walker (2001) J. Biol. Chem. 276, 8225-8230
24. V. Dolce, F. Fiermonte, M.J. Runswick, F. Palmieri and J.E. Walker (2001) Proc. Natl. Acad. Sci. USA 98, 2284-2288
25. L. Palmieri, B. Pardo, F.M. Lasorsa, A. del Arco, K. Kobayashi, M. Iijima, M.J. Runswick, J.E. Walker, T. Saheki, J. Satrustegui and F. Palmieri (2001) EMBO J. 20, 5060-5069
26. L. Palmieri, G. Agrimi, M.J. Runswick, I.M. Fearnley, F. Palmieri and J.E. Walker (2001) J. Biol. Chem. 276, 1916-1922
27. G. Fiermonte, L. Palmieri, S. Todisco, G. Agrimi, F. Palmieri and J. E. Walker (2002) J. Biol. Chem. 277, 19289-19294
28. C.M.T. Marobbio, A. Vozza, M. Harding, F. Bisaccia, F. Palmieri and J.E. Walker (2002) EMBO J. 21, 5653-5661
29. C.M.T. Marobbio, G. Agrimi, F.M. Lasorsa and F. Palmieri (2003) EMBO J. 22, 5975 5982
30. A. Vozza, E. Blanco, L. Palmieri and F. Palmieri (2004) J. Biol. Chem. 279, 20850 20857
31. G. Fiermonte, F. De Leonardis, S. Todisco, L. Palmieri, F.M. Lasorsa and F. Palmieri (2004) J. Biol. Chem. 279, 30722 30730
32. C.M.T. Marobbio, M.A. Di Noia, and F. Palmieri (2006) Biochem. J. 393, 441-446
33. S. Todisco, G. Agrimi, A. Castegna and F. Palmieri (2006) J. Biol. Chem. 281, 1524-1531
34. F. Palmieri, F. Bisaccia, L. Capobianco, V. Dolce, G. Fiermonte, V. Iacobazzi, C. Indiveri and L. Palmieri (1996) Biochim. Biophys. Acta 1275, 127-132
35. V. Stipani, A.R. Cappello, L. Daddabbo, D. Natuzzi, D.V. Miniero, I. Stipani and F. Palmieri (2001) Biochemistry 40, 15805-15810
36. B. Morozzo della Rocca, G. Lauria, F. Venerini, L. Palmieri, F. Polizio, L. Capobianco, V. Stipani, J. Pedersen, A.R. Cappello, A. Desideri, F. Palmieri (2003) Biochemistry 42, 5493-5499
37. B. Morozzo della Rocca, D.V. Miniero, G. Tasco, V. Dolce, M. Falconi, A. Ludovico, A.R. Cappello, P. Sanchez, I. Stipani, R. Casadio, A. Desideri and F. Palmieri (2005) Mol. Membr. Biol. 22, 443-452
38. Falconi M., Chillemi G., Di Marino D., D’annessa I., Morozzo della Rocca B., Palmieri L., Desideri A.
Proteins: Structure Function and Genetics 2006, in press
39. A. Palmisano, V. Zara, A. Honlinger, A. Vozza, P.J. Dekker, N. Pfanner, F. Palmieri (1998) Biochem. J. 333, 151-158
40. Truscott, K.N. and Pfanner, N. (1999) Biol. Chem. 380, 1151-1156.
41. Tokatlidis, K. and Schatz, G. (1999) J. Biol. Chem. 274, 35285-35288
42. Zara, V., Ferramosca, A. and Rassow, J. (2003b) Recent Res. Devel. Mol. Cell. Biol. 4, 101-114.
43. Zara, V., Rassow, J., Wachter, E., Tropschug, M., Palmieri, F., Neupert, W. and Pfanner, N. (1991). Eur. J. Biochem. 198, 405-410.
44. Zara, V., Palmieri, F., Mahlke, K. and Pfanner, N. (1992) J. Biol. Chem. 267, 12077-12081.
45. Zara, V., Ferramosca, A., Palmisano, I., Palmieri, F. and Rassow, J. (2003c) J. Mol. Biol. 325, 399-408.
46. Zara, V., Ferramosca, A., Papatheodorou, P., Palmieri, F. and Rassow, J. (2005) J. Cell Sci. 118, 3985-3995.
Keywords
MITOCHONDRIAL TRANSPORTERS, SITE DIRECTED MUTAGENESIS, EPR SPECTROSCOPY, PROTEIN IMPORT, MOLECULAR MODELING, TARGETING SIGNALSStructure and biogenesis of mitochondrial transport proteins
Università degli Studi di Roma "Tor Vergata"Abstract
The present project is aimed at obtaining insight into the structure, the structure-function relations and the biogenesis of the mitochondrial carriers. Through a close collaboration of three research units with specific expertise, this project will utilize a combination of biochemical and biophysical techniques aimed at clarifying the structural-dynamical-functional properties and the import mechanism of mitochondrial carriers and in particular of the oxoglutarate (OGC)and the citrate (CIC) carrier.The Bari unit will mutagenize single amino acids of mitochondrial carriers in order to identify the residues which are essential to the transport activity, the unit will also prepare plasmids and mutants (single-Cys and double-Cys mutants, mutants with a single tryptophan and a single cysteine), that will be made available to the other units participating in this project in order to carry out spectroscopic and biogenesis experiments. All the mutants will be expressed in E. coli, purified, incorporated into liposomes and assayed for transport activity. The single-Cys replacement mutants reconstituted into liposomes will also be tested for their sensitivity to permeable and impermeable SH-reagents, in the presence or absence of substrates or inhibitors. Furthermore, direct [14C]N-ethylmaleimide binding will be measured in the presence or in the absence of other SH-reagents, substrates or inhibitors. The Roma unit will perform EPR measurements of the nitroxide label >>>
Principal Investigator
Giuseppe Rotilio Università degli Studi di ROMA "Tor Vergata"Research Objectives
The secondary and tertiary structure of the mitochondrial carriers is only partially known and consequently the understanding at molecular level of their transport mechanism is still not yet defined, despite the important biological role of this class of proteins. The process of “import” of the mitochondrial carriers from the cytosol into the mitochondrion is also poorly understood.The object of this research program is to acquire structural information about the mitochondrial carriers in particular about their transmembrane domains, to study some important structure-function relations and to clarify the mechanism of the biogenesis. This aim will be pursued through a close collaboration between three research units, located in different Italian universities, that have specific expertises and that have already collaborated within each other. The results so far obtained by these units represent the starting point of the research activities of the present project, whose detailed aims for the next two years are:
- identification of structurally and functionally important amino acids in the mitochondrial transporters, particularly in the oxoglutarate carrier (OGC) ;
- localization of the important mitochondrial carrier amino acids in relation to the hydrophobic membrane core , to the hydrophilic substrate translocation pathway (channel)and to the residues involved in the substrate or inhibitor binding;
- determination of the structure of the >>>
Timescale
24 monthsNational and international background
Over one fifth of all the proteins are transmembrane proteins that cross the membrane several times with their polypeptide chain. They catalyze many important functions, one of which is the transport of molecules into and out of cells or intracellular organelles. Despite their importance, the structure of membrane proteins is known at high resolution for only few molecules and so the understanding at molecular level of their mechanism is very scanty. This situation is dependent on the fact that membrane proteins in general, and membrane transport proteins in particular, are resistant to crystallization and to the determination of high resolution structure because of their hydrophobicity and, in many cases, of their metastable nature. The elucidation of the structure and function of membrane proteins is one of the main goals of structural biology of the next decade and requires a common effort which joins together different expertises such as cloning, purification, reconstitution, spectroscopic characterization and computational modeling.The mitochondrial transporters or carriers (MCs) are a family of proteins that, with a few exceptions, are localized in the inner membrane of mitochondria (1). Their common function is to provide a link between mitochondria and cytosol by facilitating the flux of a large variety of solutes through the permeability barrier of the inner mitochondrial membrane (i.m.m.). This link is indispensable, as many physiological >>>
