- Vivi Padova
- Il BO
Responsible of the essential flow of ions and small molecules across membranes in a selective and regulated manner, allowing controlled communication with the cytosol. This consortium will bring together complementary and avantgarde approaches, specifically tailored for intracellular membranes, in order to characterize functional properties and proteostasis mechanisms (targeting to the correct membrane and turnover) of animal and plant carriers (specifically: channels and pores).
These studies require the integration of different kinds of expertise as well as the development of new methodologies, since intracellular membranes are often not accessible by canonical experimental techniques such as patch clamp. An added value of the consortium will thus be to provide common technology platforms for all participants. We aim at building a global picture of the relationships between structure, proteostasis and functon of channels and pores and using this information to design and test artificial structures.
Carriers of different intracellular compartments and semiautonomous organelles will be studied. In detail: the voltage gating anion selective channel isoforms of the outer membrane of mitochondria; the tantem pore K+ channel AtTPK3, which we recently discovered to be present in thylakoids; two glutamate transporters; three cation channels of the tonoplast; human cation channels of endosomes and lysosomes; two closely related miniature viral channels: Kesv, naturally targeted to mitochondria, and Kcv, targeted to plasma membrane through the secretory pathway.
Our research aims can be grouped as follows: 1) mechanims of channel gating and physiological role of intracellular channels 2) proteostasis (intracellular targeting, sorting and turnover) 3) role of intracellular channels in metabolism and stress response 4) molecular modeling and design of artificial channels.
In detail: 1)The functional studies will include: the identification of protein domains involved in gating; the influence of membrane lipid composition on transport properties; the relationships with calcium homeostasis; the clarification of protein interacting networks, a study that will heavily depend on proteomic facilities. 2) For what regards targeting and sorting, the signals and mechanisms for targeting and sorting will be characterized, including the relationships between oligomer formation and intracellular traffic, as well as the mechanisms for the double localization to different membranes. 3) The roles of plant pores and channels in cellular stress response and signal transduction will be established, also in view of the possible improvement of the response of crops to environmental stress. 4) molecular dynamic simulations of natural and artificial pores will be performed, in an effort to predict activity and suitability as artificial construct.
Swapping constructs among isoforms will be produced: this strategy has allowed to produce new pores with features that are intermediate with respect to those of the parent molecules; bioinformatics, biophysical and molecular approaches will be used to produce fully artificial pores.
transport carrier, plant biophysics, proteomics