Using the TOCSY, COSY and HSQC spectra, identify the spin systems and likely amino acid candidates for these spin systems.A packacge containing the NMR and mass spectral data for the oligopeptide can be downloaded here.2D- 13C-HMBC (Observe through bond couplings between 1H's and 13C for those pairs that are 2 to 4 bonds apart, but not directly bonded to one another).2D- 13C-HSQC (Observe through bond couplings between 1H's and 13C for those pairs that are directly bonded to one another).2D-ROESY (Observer though space couplings between 1H's that are separated by 2D-TOCSY (Similar to the COSY spectrum, but shows all members of a spin system for each member of a spin system).2D-COSY (Observe through bond couplings between 1H's that are separated by 2-3 bonds).Your goal is to determine both the amino acid composition and sequence for this oligopeptide. Working in your groups of 2, begin using Sparky to analyze the NMR experiments for the unknown oligopeptide in DMSO.Evaluate the LC/MS run for the unknown oligopeptide and estimate the number of amino acid residues that it contains.We are interested in discerning the different patterns that are characteristic for the different amino acids. Use iNMR to process your dipeptide spectru and share your dipeptide results with the other members of the class.Make 1H and 13C assignments and identify the dipeptide sequence.2D TOCSY with WATERGATE solvent suppression ( MLEVGPPH19SW).1D proton with WATERGATE solvent suppression ( P3913GP).Check to make certain that all material dissolve, if not the case, spin in microfuge for 5 min.Make up 10 mM samples of your unknown in 650 μL 90% H 2O/10% D 2O.Run ( zg) 2D TOCSY with WATERGATE solvent suppression ( MLEVGPPH19SW).Run ( zg) 1D proton with WATERGATE solvent suppresion ( P3913GP).Determine the offset and 90° pulse width ( paropt on p1, look for 360° null).Sit through the presentation and demonstration on using Topspin to collect 1D and 2D spectra and iNMR for processing spectra.Perform LS/MS runs on your unknown dipeptides and predict its amino acid composition from its observed mass. ![]()
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