Measurement time costs on the 700MHz Bruker Avance III NMR spectrometer

The MedInProt Grant has been spent on measurement time costs on the 700MHz Bruker Avance III NMR spectrometer equipped with 5-mm room-temperature, z-gradient TXI and 5mm Prodigy TCI H&F-C/N-D z-gradient probe-heads. A wide variety of classical and fast measurements are available for structural and dynamical characterisation of proteins. The present operating software is TopSpin 3.5 pl7.
In the frame of the present proposal four systems were studied and 2 papers were published already.

1. Structural characterization and biologyical activity of a new antimicrobial protein

Protein structure calculation necessitates high quality input data. One type of such constraints are the 1H-1H distances collected from NOESY spectra. Measurements conducted at the 700MHz spectrometer led to 976 noe restraints, and based on these the solution structure of the sfPAFB protein could be determined (with RMSD 0.3Å) .This remarkably stable 58 aminoacid residues long, cystein-rich protein shows a structure build from 5 β sheets, 4 flexible loop regions and an „abcabc” disulfide bridge pattern. Despite sequence similarity to the PAF family member is only 35%, the structures are very similar. The biological function of potential antimicrobial activity are performed by the cooperation leading partner from Innsbruck.

Detailed results are presented in the following paper:
SCIentIFIC REPOrTS | (2018) 8:1751 | DOI:10.1038/s41598-018-20002-2 1

Gyula Batta - Andrea Bodor

2. Influence of phosphorylation and interaction on the CTT-PDZ domain

We gained structural insights into the intramolecular clamp formed in the phosphorylated CTT. NMR studies were conducted for the native and phosphorylated CTT conformations and PDZ bound states.
Our results shed light also on the importance of the proline cis-/trans- configuration.

The paper appeared as Editor´s choice: The FEBS Journal 285 (2018) 46–71, 2017.

László Nyitray - Andrea Bodor

3. Lipid-protein interaction studies

By combining the NMR spectroscopy and SAXS techniques we study phospholipid bicelles and their interaction with different model peptides. We are interesed in the characterization of these systems and variation of several parameters with concentration, temperature, lipid side-chain.

Attila Bóta - Andrea Bodor

4. Characterization of modified canonical serine protease inhibitors

Our investigation is focused on SGCI and SPINK proteins and we try to elucidate how the well-chosen loop mutants affect atomic structure, and how can this be correlated with the specificity of the given protein. Primarily structural studies are conducted on unlabeled, 15N and 13C/15N proteins.

Gábor Pál - Andrea Bodor