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Laboratory of Biochemistry and Molecular Biology of Sphingolipids  

Lab. Colombo

 

Responsible 

Irma Colombo, Associate professor irma.colombo@unimi.it

 

Team 

Fausta Omodeo Salè, Associate professor fausta.omodeosale@unimi.it 

Clara Lindi, Associate professor clara.lindi@unimi.it 

Stefania Zava, Technician stefania.zava@unimi.it 

Diletta Scaccabarozzi, PhD student diletta.scaccabarozzi@unimi.it 

 

Areas of investigation 

Since several years the aim of the researches in our laboratory is the study of the quali- and quantitative profile, of the structure-function correlations and of metabolism of complex lipids, particularly phospholipids and sphingo(glyco)lipids, in animal and cellular models in both physiological and pathological conditions. 

Presently our researches are addressed to the study of: 

- alterations of phospholipid and sphingo(glyco)lipid structure and metabolism induced by Plasmodium falciparum and/or its metabolic products in tissues (liver and lung) from murine models and cell cultures (erythrocytes, endothelial cells, keratinocytes);

- alterations of the biosynthetic and catabolic metabolism of sphingo(glyco)lipids during the differentiation and/or neoplastic transformation in culture cells (mammary cells and keratinocytes);

- biochemical and molecular aspects of glycosyltransferases, in particular the human GM3 synthase (gene cloning and expression, enzyme kinetic, biochemical properties and cellular localization); 

- role of the sphingo(glycol)lipids in signalling pathways activated by Erb family and estrogen receptors in normal (HC11 and MCF-7 cells) and tumor (MDA-MB 231 cells) mammary cell lines.

 

Main techniques 

- extraction, purification and analysis of lipid components by chromatography techniques (HP-TLC, gas-cromatografia e HPLC) 

- isolation and biochemical characterization of proteins by electrophoretic (SDS-PAGE) e immunological (western blot, ELISA, immunoprecipitation) techniques, and by enzyme assays 

- purification, analysis and manipulation of plasmid, batteriophage and genomic DNA, and of RNA 

- gene cloning and expression (by transient and/or stable transfection) in prokaryotic and eukaryotic cells 

- analysis of gene structure organization, by primer extension and S1 mapping, and gene expression regulatory regions, by reporter genes 

- gene expression analysis by Northern blot, RT-PCR and RNase Protection Assay 

- analysis of DNA-protein interactions by EMSA and of protein-protein interactions by co-immunoprecipitations 

 

Selected articles 

1. F. Omodeo-Salè, L. Cortelezzi, Z. Vommaro, D. Scaccabarozzi, AM Dondorp. “Dysregulation of L-arginine metabolism and bioavailability associated to free plasma heme.” Am J Physiol Cell Physiol. (2010) 299, 148–154. 

2. S. Milani, E. Sottocornola, S. Zava, M. Galbiati, B. Berra, I. Colombo. "Gangliosides influence EGFR/ErbB2 heterodimer stability but they do not modify EGF-dependent ErbB2 phosphorylation." Biochim Biophys Acta (2010) 1801, 617-624. 

3. S. Zava, S. Milani, E. Sottocornola, B. Berra, I. Colombo. "Two active and differently N-glycosylated isoforms of human ST3Gal V are produced from the placental mRNA variant by leaky scanning mechanism." FEBS Lett (2010) 584, 1476-1480. 

4. N. Basilico, L. Cortelezzi, C. Serpellini, D. Taramelli, F. Omodeo-Salè. “Two complementary fluorimetric assays for the determination of aminoquinoline binding and uptake by human erythrocytes in vitro.” Anal Biochem (2009) 385, 371-373. 

5. F. Omodeo-Salè, L. Cortelezzi, N. Basilico, M. Casagrande, A. Sparatore, D. Taramelli. “Novel antimalarial aminoquinolines: heme binding and effects on normal or Plasmodium falciparum-parasitized human erythrocytes.” Antimicrob Agents Chemother (2009) 53, 4339-4344. 

6. S. Milani, E. Sottocornola, S. Zava, P. Berselli, B. Berra, I. Colombo. "Ganglioside GM3 is stably associated to tyrosine phosphorylated ErbB2/EGFR receptor complexes and EGFR monomers, but not to ErbB2." Biochim Biophys Acta (2007) 1771, 873-878. 

7. F. Omodeo Salè, L. Cortelezzi, E. Riva, E. Vanzulli, D. Taramelli. “Modulation of glyceraldehyde 3 phosphate dehydrogenase activity and tyr-phosphorylation of Band 3 in human erythrocytes treated with ferriprotoporphyrin IX.” Biochem Pharmacol (2007) 74, 1383-1389. 

8. E. Sottocornola, R. Misasi, V. Mattei, L. Ciarlo, R. Gradini, T. Garofano, B. Berra, I. Colombo and M. Sorice. "Role of gangliosides in the association of ErbB2 with lipid rafts in mammary epithelial HC11 cells." FEBS J. (2006) 273, 1821-1830. 

9. P. Berselli, S. Zava, E. Sottocornola, S. Milani, B. Berra, I. Colombo. "Human GM3 synthase: a new mRNA variant encodes an NH2-terminal extended form of the protein." Biochim Biophys Acta (2006) 1759, 348-358. 

10. F. Omodeo Salè, E. Vanzulli, G. Montorfano, D. Monti. “Major artefacts encountered in studying biological samples containing ferriprotoporphyrin IX.” Anal Biochem (2005) 339, 257-261.

 

Patents 

1. S. Ronchi, A. Bartorelli, B. Berra, I. Colombo. "Proteins from mammalian liver and their use in oncology." (WO 96/02567) 01.02.1996. 

2. A. Bartorelli, B. Berra, I. Colombo, S. Ronchi. "Nucleotide sequence from goat liver." (WO 97/30154) 21.08.1997. 

3. V. Naktinis, D. Concas Benevelli, B. Berra, I. Colombo, S. Ronchi, A. Bartorelli, V. A. Bumelis. "Recombinant protein and its use in therapy and diagnostics." (WO 00/63368) 26.10.2000. 

 

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