Heterotrimeric collagen peptides containing functional epitopes. Synthesis of single-stranded collagen type I peptides related to the collagenase cleavage site.
Synthetic collagen peptides containing larger numbers of Gly-Pro-Hyp repeats are difficult to purify by standard chromatographic procedures. Therefore, efficient strategies are required for the synthesis of higher molecular weight collagen-type peptides. Applying the Fmoc/tBu chemistry, a comparative analysis of the standard stepwise chain elongation procedure on solid support with the procedure based on the use of the synthons Fmoc-Gly-Pro-Hyp(tBu)-OH and Fmoc-Pro-Hyp-Gly-OH was performed. The crude products resulting from the stepwise elongation procedure and from the use of Fmoc-Gly-Pro-Hyp(tBu)-OH clearly revealed large amounts of microheterogeneities that result from incomplete imino acid acylation as well as from diketopiperazine formation with cleavage of Gly-Pro units from the growing peptide chain. Conversely, by the use of the Fmoc-Pro-Hyp-Gly-OH synthon, the quality of the crude products was significantly improved; moreover, protection of the Hyp side chain hydroxyl function is not required using the Fmoc/tBu strategy. With this optimized synthetic procedure, relatively large collagen-type peptides were obtained in satisfactory yields as highly homogeneous compounds.
Ottl
J
J
Max-Planck-Institute of Biochemistry, Martinsried, Germany.
Musiol
H J
HJ
Moroder
L
L
D000595:Amino Acid Sequence; D001671:Biochemistry / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D003094:Collagen / Q000737:chemistry*; D017364:Collagenases / Q000378:metabolism*; D000939:Epitopes; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis* / Q000276:immunology* / Q000378:metabolism
D000939:Epitopes; D010446:Peptide Fragments; D003094:Collagen; D017364:Collagenases