Mapping, characterization and specific inhibition of allosteric protein-protein interactions in signaling networks involved in tumorigenesis and Alzheimer disease

A lecture by  Péter Závodszky PhD

Péter Závodszky Bio:

Institutional Affiliation       

Research Professor, Research Centre for Natural Sciences, Institute of Enzymology,

Hungarian Academy of Sciences, Budapest;

Professor Emeritus, Department of Biological Physics, Eotves Lorand University,

Visiting Professor, Department of Chemistry and Biochemistry, UCLA, Los Angeles

Research Interest

Protein structure, signal transduction in the immune system, protein-protein interactions, protein expression and purification, allosteric regulatory mechanisms, kinase signaling pathways

Applied research

Development of the first compact analytical ultracentrifuge (Hungarian Optical Works, 1974) 

Development of acusto-optical deflector MTA SzTAKI (US and Hungarian patent 1977)

Recombinant protein expression technology (annexin, teriparatide, Richter , 2002)

Development of specific complement protease inhibitors (3 patents, 2010-16)

Educational History         

Diploma (MSc) in Nuclear Physics; Kossuth University Debrecen, Hungary

Ph.D. in Biochemistry (1971) Institute of Biochemistry, Hungarian Academy of Sciences, Budapest

Doctor of Sciences (1987) Hungarian Academy of Sciences, Budapest

Member of the European Academy of Sciences and Arts (2000)

Member of the Hungarian Academy of Sciences (2001)

Graduate Studies  

1963   Institute of Polymer Research of the Soviet Academy of Sciences, St. Petersburg, Russia

1964   Institute of Molecular Biology, Soviet Academy of Sciences, Moscow, Russia 

Postdoctoral research training  

1972-73   CALTECH, Department of Chemistry, Pasadena, CA, USA (Protein X-ray diffraction)

1975   Oersted Institutet, Copenhagen, Denmark (Protein structure research)

Research and Academic Positions       

1962-present          Research fellow, research professor, director (2007-2011), Institute of Enzymology, Research Centre for Natural Sciences   of the Hungarian Academy of Sciences                                                                                                                                                                               

1968-2009    Lecturer-professor in Biophysics, Eotvos Lorand University, Budapest

1977-78        Visiting Professor Department of Biochemistry, University of Oxford, UK 

1984               Visiting research fellow NIH, Bethesda (IgG and complement research)

1985-present          Visiting Professor, Department of Chemistry and Biochemistry, UCLA, Los Angeles,

1987-89        Visiting Professor, Abteilung für Physikalische Biochemie, Universität Regensburg

Other appointments       

Biology Division of the Hungarian Academy of Sciences, president (2008-14)

The Biochemical Committee of the Hungarian Academy of Sciences (1974-84 secretary)

Hungarian Biophysical society honorary president (president 2007-2015)

Hungarian Association for Innovation, vice president

Hungarian Foundation for Innovation, Chairman

Prime Minister’s Office, Science and technology policy advisor (1988-2002)

Hungarian Accreditation Committee, advisor (member 2005-2011)

Hungarian Stipendium Committee, President 

Associate editor for the "Archives of Biochemistry and Biophysics” (1992-2000) 

Section editor for the journal: „Immunbiology” (2011-2018)

1977-Member of Exeter College, Oxford

Hungarian Optical Works, Chairman of the Board (1999-2002)

Scientific Advisory Board member, Gedeon Richter, Pharmaceutical company (2000-2007)

Honours and Awards      

1970   Szorenyi Award

1976   Government Award for Achievements in International Scientific Cooperation 

1977   Welcome Trust Research Award

1988   German Research Council (DFG) Research Award

2003   Straub plaquette

2010   Denis Gabor award

2010   Széchenyi Prize

2013   Paladin plaquette

2015   Hungarian Cross of Merit with the Star

Publications

172 research papers, 2 books and 5 patents in the field of molecular biology and biotechnology

Abstract

Cancer and Alzheimer disease are related disorders in a way. These pathological phenomena arise as consequences of disturbances in certain cellular signaling networks. Revealing and describing the underlying protein-protein interactions in these networks is the way of understanding these signaling pathways. These pathways are organized into networks. In the case of disturbances in certain signaling pathways, intervention must be specific to the abnormal process and should not affect the normal function of the organism (side effects). The means to achieve this is to target the specific allosteric protein-protein interactions. Rassf1A is a major tumor suppressor regulating cell cycle by mitotic arrest. Down regulation of Rassf1A by specific phosphorylation by Aurora A kinase results in unregulated mitotic progression. To map the recognition signal of Aurora A on Rassf1A we designed both a pseudo-activation loop and domain deletion mutants of Rassf1A. We showed that both parts, lop and SRAH domain, are essential for the specific recognition and phosphorylation of Ser-203 of Rassf1A by Aurora A.

The studying of interaction networks of ROCK2 was further broadened into the direction of neurodegenerative disorders, in particular Alzheimer’s disease. ROCK2 was shown to phosphorylate Amyloid Precursor Protein (APP) and β-secretase (BACE1) on their intracellular flanking regions. Inhibition of this phosphorylation event reduces the formation of Aβ40 and Aβ42, both relevant peptides in Alzheimer disease. Since ROCK2 has several other, physiologically essential downstream targets, a protein-protein interaction (PPI) inhibitor could block amyloid formation selectively. Bioinformatic and physical methods were used to map the interactions between ROCK2 and its target proteins. We developed a screening cascade to find pathway specific inhibitors for the ROCK2 versus APP or BACE interaction. We also showed that APP binds to ROCK2 only in the presence of BACE1, indicating that binding of BACE1 induces a conformation change on ROCK2 and the newly formed structure is capable of interacting with APP. This result implies that ternary interaction is required for the proper phosphorylation of APP which is required for the trafficking of APP to the early endosome.We have revealed that a potentially allosteric binding site exists for the APP-ROCK2 interaction. To locate this binding site on ROCK2, we designed and expressed several mutants. Our results show that allosteric communication between the ATP and substrate binding sites does exist indeed. A ROCK2 inhibitor could have an anti-Alzheimer’s disease potential. We tested a library of small-molecule putative PPI inhibitors both for ROCK2 binding using DSF and for PPI inhibition using a fluorescence polarization assay. We identified 11 potential pathway-specific allosteric drug candidates.