NMR Analysis of Dog Erythrocytic Membrane Antigen

Aim: NMR spectroscopy analysis of dog erythrocytic membrane antigen in order to differentiate blood groups in dogs

Methodology: The purified dog erythrocytic membrane glycoprotein of DEA1.1 positive and negative blood were subjected in to NMR spectroscopy analysis. One-dimensional C13-NMR spectra were acquired at 25°C on a high resolution spectrometer Fourier 300 MHz (Bruker’s, (USA) using the first increment of the pulse sequence NOESY-presat, 128 scans, sweep window 20 ppm, 32 k points and relaxation delay 5 seconds. The spectra were processed and analyzed with Bruker’s. TopSpin software 300 zero-filing to 64 k points and line broadening 0.5 Hz22 and MestReNova 8.1 software (Mestrelab Research, Santiago de Compostela, Spain).

Results: The DEA 1.1 positive and negative membrane glycoprotein showed chemical shift with minimum spectral difference and functional group, CH3CO at the level of 20-30 ppm, RCH2Cl at the level of 35-45 ppm, RCH2OH, C=O (in acids and esters) at the level of 170 ppm were identified in both DEA 1.1 positive and negative membrane glycoprotein in NMR analysis of canine erythrocytic membrane antigen.

Conclusion: NMR studies of conformational changes of membrane proteins in response to small molecule and protein ligands and the changing lipid environment at different physiological states of cellular membranes. The exploration of the structural and mechanistic biology of membrane proteins by NMR has a bright future and bring many new exciting discoveries.