1H NMR analysis and in vitro bioactivity of Leu27-bGRF(1-29)NH2 and its D-Ala2 and des-(Tyr1-Ala2)-analogs.
Relative growth hormone-releasing potencies of bovine growth hormone-releasing factor (bGRF) analogs bGRF(1-44)NH2 (I), Leu27-bGRF(1-29)NH2 (II) and D-Ala2, Leu27-bGRF(1-29)NH2 (III) in in vitro bovine anterior pituitary cell cultures were determined to be 100%, 48% and 77%, respectively. The potencies of II and III, although numerically different, were not statistically different. Leu27-bGRF(3-29)NH2 (IV) was approximately 10,000 times less potent than 1. 1H NMR studies of peptides II, III and IV in 35% d3-2,2,2-trifluorethanol (TFE)/65% phosphate buffer at pH 4 revealed very similar, highly helical secondary structures in the 8-29 region, with only subtle differences at the N-termini. This lack of correlation between secondary structure in solution and in vitro bioactivity suggests that either 1) the biological conformations induced at the GRF receptor for II and III vs. IV are different from those generated in TFE/buffer, 2) similar secondary structures may be necessary but not sufficient for the observed bioactivity or 3) residues 1 and 2 of analogs II and III are important contact residues crucial for effective GRF-receptor interaction.
Kloosterman
D A
DA
Physical and Analytical Chemistry Research, Upjohn Company, Kalamazoo, MI 49001.
Scahill
T A
TA
Hillman
R M
RM
Cleary
D L
DL
Kubiak
T M
TM
D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D002417:Cattle; D002478:Cells, Cultured; D013006:Growth Hormone / Q000378:metabolism*; D013007:Growth Hormone-Releasing Hormone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000494:pharmacology; D010902:Pituitary Gland / Q000166:cytology / Q000187:drug effects*; D011487:Protein Conformation; D011522:Protons; D017337:Sermorelin / Q000031:analogs & derivatives*; D013329:Structure-Activity Relationship
D010446:Peptide Fragments; D011522:Protons; C074709:somatotropin releasing hormone (1-29)NH2, Leu(27)-Ala(2)-; C074710:somatotropin releasing hormone (3-29)NH2, Leu(27)-; C061235:somatotropin releasing hormone (1-29) amide, 27-Leu-; D017337:Sermorelin; D013006:Growth Hormone; D013007:Growth Hormone-Releasing Hormone