Stefano Minguzzi

Name

Stefano Minguzzi

Name: Stefano Minguzzi
Nationality: Italian

Intana Bioscience GmbH
Lochhamer Str. 29a
D-82152 Planegg/Martinsried

Contact details: T: +49 (0)89 89 55 72 84
E mail: stefano.minguzzi@intana.de

Biosketch

Stefano Minguzzi achieved both Bachelor and Master degrees in Molecular Biology at the University of Bologna. He carried out his master thesis for 1 year in the department of Microbiology of Bologna. The title of the thesis was “Studies on the butane metabolism of Rhodococcus sp. BCP1 expression of AlkB gene and type of oxidation used." In 2009, was granted with an Irish Research Council scholarship to undertake a PhD program in the Nutritional Genomics lab in Dublin City University (Ireland). Under the supervision of Dr. Anne Parle-McDermott, he worked on the human gene MTHFD1L involved in the folate metabolism. Disease association studies have been carried out on this gene and microRNA regulation has been also studied. Stefano joined Intana Bioscience GmbH (Munich, Germany) as a Marie Curie postdoctoral fellow on the 15th of May 2012. His Sphingonet project “Linking proteins and their binding partners to lipid nanodiscs” will be carried out mainly at Intana

 

Project title: Linking proteins and their binding partners to lipid nanodiscs

Project summary:

Many interactions between cellular biomolecules take place at the cellular membrane, but in vitro assays, mimicking membrane bound interactions have been complicated to establish. The nanodisc technology is an emerging artificial membrane system for the study of membrane proteins. Being more stable and monodisperse than conventional model membranes such as liposomes, bicelles or micelles, the nanodisc is an excellent alternative for single-molecule fluorescence applications. The aim of this project is to develop a method based on linkers for attaching proteins and their binding partners to lipid nanodiscs. Protein interactions will be then studied using FCCS (Fluorescence Cross Correlation Spectroscopy) based method at Intana Bioscience GmBH. Click chemistry will be employed to produce linkers featuring azido or tetrazine moieties at SiChem. To increase residence time and decrease motility in the nanodisc, photo-cross linkable lipids such as diazirines and their covalent coupling to other lipids in the disk might be used. Proteins of interest will be equipped with an artificial amino acid providing a strained cyclooctyne or cyclooctene group (Plass et al., 2011, 2012). Proteins will be expressed in E. coli and purified at EMBL. Alternatively, SNAP‐, CLIP‐, or HALO‐ tags will be used to attach proteins to nanodiscs featuring the respective benzylguanine etc lipid linkers. Rasterscan Image Correlation Spectroscopy (RISC) and Number and Brightness (N&B) analysis will be also used. Our approach is focused on membrane proteins having high potential for future pharmaceutical development such as receptor tyrosin kinases (RTKs), G-protein coupled receptors (GPCRs) some of which are involved in sphingolipid homeostasis. For example, Sphingosine-1-phsophate is a bioactive lipid mediator that is critical in many biological processes (cell survival, mobility, proliferation, etc) and acts through five GPCRs (S1P1-5). Sphingosylphosphorylcholine (SPC) is implicated in a number of physiological and pathological processes as well as has high-affinity with several GPCRs such as G protein-coupled receptor 4 (GPR4), Ovarian cancer G protein-coupled receptor 1 (OGR1) or G protein-coupled receptor 12 (GPR12). By adapting the FCCS and RISC approach to those sphingolipid receptors, we will provide a new method for determining protein interactions at the cellular membrane

Supervisors

Dr. Stefan Hannus (Intana Bioscience GmbH, Munich, Germany)

Dr. Carsten Schultz (EMBL Heidelberg, Germany)

Biosketch 1st scientific supervisor:

JDr. Stefan Hannus studied Biology at the University Regensburg with focus on molecular biology and biochemistry. Before starting his PhD thesis, he joined the Lab of Prof. Dr. E. Hurt in Heidelberg investigating ribosomal transport across the nuclear pore complex. He received his predoctoral education at the EMBL in Heidelberg and the Max-Planck-Institute for Biochemistry in Martinsried conducting research on the biogenesis of ribonuclear protein complexes. In 2001 Stefan Hannus started as scientist at GPC Biotech AG where he developed and established FCCS as tool to study compound target interactions. In 2008 he founded jointly with Dr. Frank Becker Intana Bioscience GmbH