Svetla Stoilova-McPhie, Ph.D.
Svetla Stoilova-McPhie, Ph.D.
Molecular Biology and Biophysics, University of Sofia,
Doctor of Philosophy (Physics) Hungarian Academy of Sciences, 1992
Post-Doctoral (Cryo-electron microscopy and biophysics)
Oxford University and University of Warwick (UK), 2003-2008
Wadsworth Center NY State Department of Health, 2001-2003
University of Leeds (UK), 1995-2001
IGBMC, Strasbourg (France), 1992-1995
About the Lab
Structure and Function of Biological Macromolecules
Cryo-electron microscopy (Cryo-EM) and crystallography is a powerful method to study the structure, assembly and dynamics of biological macromolecules and their interactions in close to native conditions and subnanometer resolution. This approach is straight forward for three-dimensional (3D) reconstruction of soluble complexes with a mass above 500kDa and known oligomeric structure, as viruses and cage-like complexes [Zhou ZH. 2008, Curr Opinion Str. Biol. 18:281-228]. Cryo-EM is capable of visualizing fully hydrated lipid structures, as lipid bilayer and phases at atomic resolution. Thus protein-protein and protein-membrane interactions can be followed at a molecular level and their structure resolved in close to native environment [Fujiyoshi Y. and Unwin N. 2008, Curr Opinion Str. Biol. 18:587-592].
In our lab we focus on solving the structure and macromolecular organization of biological complexes in their native environment by Cryo-EM to fully understand their function in physiological and pathological conditions.
1. Our main project is to solve the structure of membrane-bound coagulation factors VIII and V and their complexes, combining Cryo-EM with biophysical and molecular biology techniques.
Normal hemostasis is achieved through a delicate balance between precoagulant and anticoagulant proteins, known as coagulation factors, which interact with each other and cells to ensure the formation of a non-pathological blood clot at the site of vascular injury. In the center of the blood coagulation is the formation of two membrane-bound complexes: the Tenase and the Prothrombinase complexes onto the activated Platelet surface. The successful formation of these two complexes ensures the extraordinary special and temporal regulation of the coagulation process. Factor VIII and Factor V are the co-factors in the Tenase and Prothrombinas complex, respectively. Binding of the co-factors to the respective proteases onto the negatively charged membrane surface increases their proteolytic activities 200,000 to 300,000 times, leading to the efficient generation of Thrombin and the blood clot.
2. Nanotechnology designing lipid nanotubes and vesicles capable of carrying active biological complexes.
3. Cryo-electron tomography of whole cells.
Parmenter C. and S. Stoilova-McPhie 2008. Binding of recombinant human coagulation factor VIII to lipid nanotubes. FEBS Letters, 28:1657-60.
Parmenter C.D.J., M.C. Cane, R. Zhang and S. Stoilova-McPhie 2008. Cryo-electron microscopy of coagulation Factor VIII bound to lipid nanotubes. Biochem Biophys Res Comm 366, 288-293.
Stoilova-McPhie S., C.D.J. Parmenter, K. Segers, B.O. Villoutreix, G.A.F. Nicoalaes 2008. Defining the structure of membrane-bound human blood coagulation Factor Va. J Throb and Haemost 6, 76-82.
Clokie M.R.J., K. Thalassinos, P. Boulanger, S.E. Slade, S. Stoilova-McPhie, M.C. Cane, J.H. Scrivens and N.H. Mann 2008. A proteomic approach to the identification of the major virion structural proteins of the marine cyanomyovirus S-PM2. Microbiology 154:1775-82
Stoilova-McPhie S., B.O. Villoutreix, K. Mertens, G. Kemball-Cook, A. Holzenburg 2002. 3-dimensional structure of membrane-bound coagulation Factor VIII: modeling of the Factor VIII heterodimer within a 3-dimensional density map derived by electron crystallography. Blood 99: 1215-1223.
Lee SC, S. Stoilova-McPhie , L. Baxter , V. Fulop , J. Henderson , A. Rodger , D.I. Roper , D.J. Scott , C.J. Smith , J.A. Morgan 2007. Structural Characterisation of the Insecticidal Toxin XptA1, Reveals a 1.15 MDa Tetramer with a Cage-like Structure. J Mo. Biol 366: 1558-68.
Simidjiev I, S. Stoylova, H. Amenitsch, T. Javorfi, L. Mustardy, P. Laggner, A. Holzenburg and G. Garab 2000. Spontaneous formation of membranes from non-bilayer lipids and antenna complexes of plant thylakoids. Proc Natl Acad Sci 97:1473-6.
Stoylova S.S., P.J. Lenting, G. Kemball-Cook and A. Holzengburg 1999. Electron crystallography of human blood coagulation factor VIII bound to phospholipid monolayers. J. Biol Chem 274:36573-8.