Abstract: Two-Dimensional NMR methods were used to assign methyl and methylene group resonances in the NMR spectra of 2-Zn insulin. 10 methyl group of Ile, Leu, Val,Thr resonances are specifically assigned using nuclear overhauser effect (NOESY) spectra of insulin in combination with the information of X-ray structure.
Assignment of other group resonances were based on the analysis of Two-Dimensional spectra of the J-Correlated (COSY).
Insulin exists in the monomeric state at pH 11.0, where spin diffusion is not a serious problem.

Key words: 2-D NMR spectra, Insulin, Assignment of resonances