The Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) is the most important independent research funding organisation in Germany. It supports both scientific and humanistic studies and aims to promote top-level research in various disciplines at universities and research institutions. The main focus is on funding projects proposed by the scientific community, with an emphasis on knowledge-driven research.
Regulated phosphorylation on specific serine, threonine or tyrosine residues of proteins is an important control mechanism for processes as diverse as cell growth and proliferation, differentiation, metabolism, apoptosis and immune response. Approximately one third of all eukaryotic gene products can be post-translationally phosphorylated.
Consequently, a growing body of data shows that phosphorylation is directly or indirectly involved in virtually all signalling processes. Important physiological processes such as learning and forgetting, which are the subject of intense research, have been shown to be are also under the control of specific phosphorylation.
Head of Department
Prof. Dr. Stefan Szedlacsek is the Head of the Enzymology Department at the Institute of Biochemistry of the Romanian Academy. He holds a PhD degree in Biotechnology from Polytechnical University of Bucharest as well as a MSc in Organic Synthesis (Polytechnical University, Bucharest) and MSc in Mathematics (University of Bucharest). As a visiting scientist, he performed research in the field of cholesterol metabolism at the University of Illinois at Urbana-Champaign (USA), where he succeeded to evidence a new pathway in the metabolism of oxysterols. He is an “Alexander von Humboldt” fellow and worked in Germany, in the Institute of Biochem More...
The main objectives of the project are the following:
1. Kinetic evaluation of the influence of Kinase Interaction Motif (KIM) on the enzymatic activity of PTP-SL
2. Kinetic evaluation of the influence of phosphorylation on Thr 253 of PTP-SL on the intrinsic phosphatase activity.
3. Crystallization of PTP-SL containing the KIM region and (in case of successful crystallizatron) solving the corresponding 3D-structure
4. Crystallization of the complex between PTP-SL containing the KIM region and ERK2 and (in case of successful crystallization) solving the corresponding 3D-structure.
|
Step |
Key Activities/Results |
Observations |
|
Cloning and purification |
Generation and purification of truncated and mutant forms of PTP-SL, ERK2 and MEK |
Obtained high purity proteins |
|
Complex formation |
Formation of ERK2-PTP-SL complexes |
Problems in separating phosphorylated forms of ERK2 |
|
Structural Analysis and Modelling |
Cross-linking and modelling to predict interactions |
Crystallisation of complexes not achieved |
|
Enzyme Kinetics |
Determination of kinetic parameters of PTP-SL constructs |
Activating effect of KIM region limited by complex dissociation challenges |
Published papers:
1. „Identification and specificity profiling of protein prenyltransferase inhibitors using new fluorescent phosphoisoprenoids”: Dursina B, Reents R, Delon C, Wu Y, Kulharia M, Thutewohl M, Veligodsky A, Kalinin A, Evstifeev V, Ciobanu D, Szedlacsek SE, Waldmann H, Goody RS, Alexandrov K; JACS; 128, 2822-35; 2006; https://pubmed.ncbi.nlm.nih.gov/16506760/
2. „Interface Analysis of the Complex between ERK2 and PTP-SL”; Balasu MC, Spiridon LN, Miron S, Craescu CT, Scheidig AJ, Petrescu AJ, Szedlacsek SE.; PLoS One; 4(5); e5432; 2009 https://pubmed.ncbi.nlm.nih.gov/19424502/
Fellowship/Scholarship
PhD Student: Badea Rodica
2003 Fellowship (three months) at the Department of Structural and Molecular Biophysics, Max Planck Institute, Dortmund, Germany;
2004 Scholarship (6 weeks) at the Department of Structural and Molecular Biophysics of the Max Planck Institute, Dortmund, Germany;
Romanian Academy