Nmr studies of human cystatin C - stable isotope labeling of human cystatin C

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