13C-nuclear magnetic resonance study of [85% 13C-enriched proline]thyrotropin releasing factor: 13C-13C vicinal coupling constants and conformation of the proline residue.
To understand fully interactions between peptides and cellular receptors, peptide side chain conformation must be defined. In many cases the complexity of proton nuclear magnetic resonance (NMR) prevents this but the present work demonstrates this problem can be solved by using 13C enrichment. Selective 13C enrichment of a natural peptide hormone has been achieved by preparing [85% 13C-enriched proline]thyrotropin releasing factor which was examined by 13C NMR spectroscopy at various pH values. Because of the 13C enrichment, one-bonded and three-bonded (vicinal) 13C-13C coupling constants have been determined. The latter vary from 0 to 5 Hz and show bond angle dependence. These data indicate that in this hormone the pyrrolidine ring is not free but fixed in the Cgamma-endo puckered conformation. It has also been possible to assign chemical shift values for a second order 13C NMR spectrum.
D002247:Carbon Isotopes; D005998:Glycine / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy; D009842:Oligopeptides; D011392:Proline / Q000032:analysis; D011487:Protein Conformation; D013973:Thyrotropin-Releasing Hormone
D002247:Carbon Isotopes; D009842:Oligopeptides; D013973:Thyrotropin-Releasing Hormone; D011392:Proline; D005998:Glycine