STUDY OF THE TRANSMEMBRANE PROTEIN CD79B BY MULTIDIMENSIONAL PULSE NMR SPECTROSCOPY
Abstract
In the present work, using the cell-free expression system, we prepared the proteins CD79A/CD79B labeled by stable isotopes of carbon-13 and nitrogen-15. It is shown that target proteins are mostly localized in the pellet of the cell-free expression system, which is consistent with their membrane nature and the presence of a transmembrane domain in their structure. Physicochemical parameters of the CD79A/CD79B samples were defined to obtain the multidimensional correlation NMR spectra of high resolution. The analysis of the obtained correlation spectra shows that under experimental conditions, CD79B exists in the disordered state. The splitting of the signal from the NH-group of the side chain of single tryptophan residue indicates the presence of slow conformational transitions in this region of the polypeptide chain. The addition of the trifluoroacetic acid to the solution of CD79B in DMSO leads to the destruction of the intermolecular bonds of “protein micelles” and the formation of its monomeric form that is well detectable by NMR.
About the Authors
E. V. Pankratova
Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus, Minsk
Belarus
Junior researcher
Belarus
Junior researcher
V. V. Britikov
Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus, Minsk
Belarus
Researcher
Belarus
Researcher
S. A. Usanov
Institute of Bioorganic Chemistry of the National Academy of Sciences of Belarus, Minsk
Belarus
Corresponding Member, D. Sc. (Chemistry), Professor
Belarus
Corresponding Member, D. Sc. (Chemistry), Professor
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