Proton NMR and photochemically induced dynamic nuclear polarization studies of peptide fragments obtained by controlled proteolysis of mouse epidermal growth factor.
Controlled proteolysis of epidermal growth factor from the mouse leads to fragments of mouse epidermal growth factor containing residues 1-48 and 1-45. The COOH-terminal pentapeptide appears to play a crucial role in determining the hydrophobic interactions between the hormone and the stationary phase during gel chromatography on TSK-125 gel. Proton NMR studies indicate that the overall structure of mouse epidermal growth factor is retained in the protein devoid of the COOH-terminal pentapeptide, while subsequent cleavage of the peptide bond between Arg-45 and Asp-46 starts to perturb the proton resonances most characteristic of the tertiary structure of the hormone, especially those from the aromatic ring protons of Tyr-37. Consequently, photochemically induced dynamic nuclear polarization experiments show an increased exposure of Tyr-37 in the fragment of mouse epidermal growth factor containing residues 1-48. Nuclear Overhauser data suggest that structural changes do occur on fragmentation but seem to be localized in the tiered-beta-sheet domain which contains Tyr-37.
D000595:Amino Acid Sequence; D000818:Animals; D004815:Epidermal Growth Factor / Q000302:isolation & purification / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008297:Male; D051379:Mice; D010446:Peptide Fragments / Q000302:isolation & purification; D010777:Photochemistry; D011487:Protein Conformation; D013363:Submandibular Gland; D014357:Trypsin / Q000378:metabolism
D010446:Peptide Fragments; D004815:Epidermal Growth Factor; D014357:Trypsin