NMR STUDIES OF HUMAN CYSTATIN C – STABLE ISOTOPE LABELING OF HUMAN CYSTATIN C

Abstract

The objective of this study was to obtain a double and triple labeled human cystatin C (hCC). Another objective was to record sets of 2D and 3D NMR spectra of the hCC dimer (in a solution) using a 700MHz spectrometer. The data obtained during attempts to determine the NMR structure should provide useful information about chemical shifts of amino acid residues. They will certainly accelerate solving the human cystatin C NMR structure. In this paper the main focus is put on triple isotopic labeling, protein overproduction and NMR analysis of hCC. The first two processes lead to obtaining hCC labeled with stable isotopes of carbon (¹³C), nitrogen (¹⁵N) and hydrogen (²H) (double labeled hCC was obtained with a similar method). The obtained protein was later used for the purpose of NMR spectra.