NMR Structural characterisation, sito-directed mutagenesis and binding properties of structurally homologous proteins belonging to the lipocalin family: a tool for the investigation of a common folding mechanism and functional properties.

Starting date
November 5, 2002
Duration (months)
24
Departments
Biotechnology
Managers or local contacts
Molinari Henriette
Keyword
Molecular and structural biology, Site-directed mutagenesis, Nuclear magnetic resonance, Lipocalins, Folding, Binding

The task of this research unit in the framework of this project will be to perform a structural characterisation and a detailed study of the folding and binding mechanisms of structurally related proteins belonging to the lipocalin family.
We will initially concentrate on bovine and porcine beta-lactoglobulins. These proetins share high sequence identity (> 60%)and similarity (> 80%) but differ in the folding and binding properties. The comparison of their physico-chemical properties together with the anlysis of specifically designed mutants and to the study of lactoglobulins belonging to other species, will allow us to investigate the determinants of stability, folding and binding, thus enabling the elucidation of functional properties.

The main tasks of our research unit can be summarised as follows:
  1. structural characterisation of recombinant porcine beta Lactoglobulin (PLG)enriched in 15N and doubly enriched (in 13C and 15N);
  2. comparative folding studies of bovine beta lactoglobulin (BLG) and PLG and characterisation of the denatured state. This study will be performed also on mutants;
  3. conformational analysis of BLG and PLG fragments;
  4. binding studies of BLG and PLG with fatty acids with different chain lenght;
  5. modelling and thermodynamic studies.

Sponsors:

Ministero dell'Istruzione dell'Università e della Ricerca
Funds: assigned and managed by an external body
Syllabus: FIRB

Project participants

Federico Fogolari
Henriette Molinari
Raffaella Ugolini

Activities

Research facilities