Synthesis and physical properties of (hydroxyproline-proline-glycine) 10: Hydroxyproline in the X-position decreases the melting temperature of the collagen triple helix

• Published in: Archives of biochemistry and biophysics Vol. 219; no. 1; pp. 198 - 203
• Main Authors: Inouye, Katsuhiko, Kobayashi, Yuji, Kyogoku, Yoshimasa, Kishida, Yasuo, Sakakibara, Shumpei, Prockop, Darwin J.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.11.1982
• Subjects: Collagen; Drug Stability; Hydroxyproline; Kinetics; Peptides - chemical synthesis; Protein Conformation; Protein Denaturation; Temperature
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1016/0003-9861(82)90149-7

The collagen-like polytripeptide (hydroxyproline-proline-glycine) 10 was synthesized with a solid-phase procedure. Analytical ultracentrifugation indicated that the peptide in aqueous solution at 6 °C had a molecular weight of 2550, the expected size of a single chain. The peptide had a relatively small negative optical rotation at 578 nm, and it did not show a thermal transition as is seen with collagen or collagen-like polytripeptides which form triple helices. At low temperatures in aqueous solution, the circular dichroism spectrum was similar to that of triple-helical collagen and collagen-like peptides in that there was a positive peak at 224 nm and a negative peak at 200 nm. The amplitudes of the peaks, however, were considerably less than the peaks obtained with triple-helix proteins and peptides. Since (proline-proline-glycine) 10 was triple helical under the same conditions, the results demonstrated that hydroxyproline in the X-position of the repeating -glycine- X-Y- sequences decreases rather than increases, the thermal stability of the triple helix. This positional specificity cannot be explained by any of the current models for the structure of the triple helix or any of the current proposals for how hydroxyproline stabilizes the structure.