11.1. 2016, Proteinové struktury

PhD Project1:

Study of allosteric communication in chimeric two-domain proteins upon binding to TRP channels

 

PhD Project2:

Chimera two-domain proteins containing PDZ3 and their interactions

 

Research topics:

Nowadays, the studies of protein structures move from mapping of their basic function to studies of communications between their elementary units – protein domains. The information about concerted domains behaviour can be used to decipher inter/intra domain communication, their allosteric modulation and interactions facilitating their functions. Such phenomena belong to the most important and interesting issues explored recently.

The proposed projects are focused on new synthetic proteins composed from two functional different protein domains. We intend to study them in a wide complexity – by bioinformatics analysis, structural and in vivo studies revealing their coupled behaviour (e.g. allosteric modulation) up to biotechnology/ medicine applications. The studied system in question is based on intracellular protein domains called PDZ (from ZO-1, INAD) serving as cytoskeleton and membrane-proteins anchors linked to artificial protein molecules. These fusion synthetic chimeras will be studied upon their ability to bind transient receptor potential (TRP) channels as their natural occurring binding partners. Through identified interactions with the binding partner we will understand two-domain proteins behaviour in cell signalling processes and activation/inhibition of TRP ion receptors that are involved in ion cell homeostasis.

Aims: Expression and purification of recombinant proteins of two-domain fusion proteins (containing PDZ) and parts of TRP channels using chromatography techniques. Biochemical, biophysical, structural and in vivo characterizations of the proteins and studies their interactions and allosteric modulations with their native intracellular binding partners.

Methods: molecular biology techniques (cloning, mutagenesis), proteins purification / fplc, fluorescence spectroscopy, thermal shift assay, dynamic light scattering, circular dichroism, NMR method / X-ray studies, in vivo studies in human tissue cultures, bioinformatics studies (molecular modeling, ligand docking, etc.), applied science: coating of biotechnologically/medicinally functional two-domain proteins on 2D nano-layers

It´s prein cooperation with Institute of Organic Chemistry and Biochemistry CAS

Supervisor: Ing. Jan Teisinger, Ph.D.