PI: Research description | Bossi
Elena Bossi
Affiliation
Department of Biotechnology and Life Sciences
University of Insubria,
Via J.H. Dunant 3,Varese Italia
E-Mail: elena.bossi@uninsubria.it
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Target
Structure-function relationships of membrane proteins involved into the reuptake of neurotransmitters, nutrient and divalent cation (SLC6, SLC11, SLC15).
These proteins are studied using electrophysiology, molecular biology immunochemistry, HPLC and optical techniques. The main goals are the functional characterization, the pharmacology and the identification of the structural determinants of ion and substrate selectivity.
The molecular mechanism of substrate translocation is mainly investigated with biophysical approach, studying the different kinds of current elicited by the transporters.
The modulation of transporters activity by extracellular signals and second messenger pathways is also addressed.
Recently the nanoparticle membrane crossing was studied in Xenopus laevis oocytes by investigating biophysical parameters coupled to fluorescent approach (SOFA, single oocytes fluorescence assay).
Method
The research activity involves the use of several state-of the art methodologies, including heterologous expression in oocytes and in cell lines, patch-clamp, voltage-clamp, confocal microscopy, fluorescence resonance energy transfer, caged–compounds photolysis, fluorescent ion indicators, green fluorescent protein constructs, site-directed mutagenesis, immunochemistry (SOC, western Blot and immunohistochemistry) as well as other techniques in molecular biology and cell culture. Quantitative biophysical analysis of the different transport currents aims to obtain detailed description of transporter function and transport cycle steps.
Main areas of research
- Functional characterization of membrane transporters wild type and mutants
- Transport cycle: substrate transport at the molecular level
- Transporter interactions with substrates and inhibitors
