Expression and purification of metalloproteins for structural studies and
Paramagnetic restraints in hybrid methods for protein structural analysis
11th-13th May 2016, Sesto Fiorentino, Florence, Italy
Registration deadline : 15th April 2016
No fee is requested for pNMR funded fellows.
Registration is opened to external participants (in limited number), but involving a registration fee (140€). Please contact us for more information.
Lunches and dinners included. Travel and accommodation borne by participants.
Content:
| The course will cover the practical aspects of : 1) recombinant expression of metalloproteins and strategies for the assembly of metallocofactors, 2) protein purification and preparation of a solution NMR sample, 3) acquisition of solution NMR spectra of paramagnetic proteins for the measurement of pseudocontact shifts and residual dipolar couplings, 4) analysis of the paramagnetic restraints for the structural characterization of proteins, 5) NMR/X-ray hybrid structural refinement. Attendees will gain hands-on experience on the expression and preparation of an NMR sample in a wet lab, the setup of NMR experiments, the use of software tools like FANTEN, for the analysis of the paramagnetic restraints, and REFMAC-NMR, for the protein structure refinement through X-ray and paramagnetic NMR restraints |
Location
This Applied Training Course will take place at the CERM, Via Sacconi 6, 50019 Sesto Fiorentino (FI), Italy, on the scientific campus (Polo Universitario) of the University of Florence.
It is organised by Dr. Giacomo Parigi of CIRMMP, and Prof. Claudio Luchinat, from Giotto Biotech, one of our industrial partner.
Accommodation
Accommodation is not provided. A possible accommodation in Sesto Fiorentino is Hotel Villa Stanley http://www.villastanley.it/ Please, confirm your participation and possibly provide us the details of your accommodation asap.
Program
The course will take place from 9.30 to 18.00
Program
| Wednesday 11th May | |
| 9.30-9.40 | Welcome |
| 9.40-11.15 | GiottoBiotech staff |
| Recombinant protein expression: how to approach | |
| 11.15-11.30 | Coffee Break |
| 11.30-13.00 | GiottoBiotech staff |
| Extraction of cytochrome c from E. coli pellet | |
| 13.00-14.30 | Lunch |
| 14.30-18.00 | GiottoBiotech staff |
| Extraction of cytochrome c from E. coli pellet | |
| Thursday 12th May | |
| 9:30-11:00 | M. Fragai |
| Strategies for metal exchange in metalloproteins | |
| 11.00-11.15 | coffee break |
| 11.15-13.00 | GiottoBiotech staff |
| Purification of cytochrome c | |
| 13.00-14.30 | Lunch |
| 14.30-18.00 | E. Ravera |
| NMR of paramagnetic molecules with practicals | |
| Friday 13th May | |
| 9.30-10.45 | V. Calderone |
| From protein crystals to structural refinement: a quick overview | |
| 10.45-11.00 | coffee break |
| 11.00-12.00 | G. Parigi |
| Paramagnetic restraints for protein structure calculations | |
| 12.00-13.00 | A. Carlon |
| Practicals on FANTEN and REFMAC-NMR | |
| 13.00-14.30 | Lunch |
| 14.30-16.30 | A. Carlon |
| Practicals on FANTEN and REFMAC-NMR | |
| 16.30-17.30 | C. Luchinat |
| How to tackle protein structural data from solution and solid state : An integrated approach |
Registration
More information coming soon! 🙂
Last Update 12/01/2016.