(Prolyl-hydroxylprolyl-glycine)10

Glycine is found as every third residue along the entire length of triple helices in fibrillar collagens, but the triple-helix regions of nonfibrillar collagens and other proteins usually contain one or more interruptions in this repeating pattern. A set of four peptides was designed to model the effect of interruptions in the (Gly-X-Y)n repeating pattern on triple-helix formation, stability, and folding. Into the middle of the stable triple-helical peptide (Pro-Hyp-Gly)10, an interruption was introduced representing one of the four possible categories: a glycine deletion, a deletion of a hydroxyproline (Y position), an alanine insertion, or a glycine to alanine substitution. As shown by sedimentation equilibrium, NMR, and CD studies, the introduction of an interruption still allowed formation of trimers in solution, but with marked decrease in stability. The degree of destabilization and the thermodynamic basis for the loss of stability depended on the type of interruption. The glycine substitution and alanine insertion were the least disruptive, followed by the hydroxyproline deletion, with the glycine deletion being the most destabilizing. Our results suggest that the breaks in these peptides affect both the triple-helical conformation and the monomer conformation. These studies provide a basis for considering the structural and functional consequences of different kinds of interruptions in collagen.

A novel conjugate, PHG10 dye, was synthesized using a collagen peptide and a near-infrared (NIR)-responsive dye to achieve targeted cytotoxicity. The collagen peptide motif, -(Pro-Hyp-Gly)10 - (PHG10), was incorporated for targeting collagen fibrils that are excessively produced by activated fibroblasts around tumor cells. PHG10 dye was purified by HPLC and identified by MALDI-MS. The phototoxicity and cytotoxicity of PHG10 dye were examined using human glioma cells (HGCs). Fluorescent images indicated that PHG10 dye preferably assembled to collagen-coated HGCs compared with noncoated HGCs. Under irradiation with NIR light, effective cytotoxicity was observed on collagen-coated HGCs within 20 min. Because phototoxicity and cytotoxicity are dependent on the assembled amount of PHG10 dye, the targeting of collagen fibrils by the collagen peptide motif PHG10 is assured.

Collagen peptidomimetics have been synthesized as an alternative to natural collagen. The incorporation of unnatural residues such as peptoids in the collagen sequences can demonstrate potent and specific biological activity and enhance the biostability against enzymatic degradation. Furthermore, the use of achiral peptoids simplifies synthetic strategies by reducing racemization problems. The peptoid residue N-isobutylglycine (Nleu) has been successfully incorporated into a series of collagen mimetics composed of Gly-Pro-Nleu, Gly-Nleu-Pro, and Gly-Nleu-Nleu. The discovery of template-assembled collagen mimetics and metal binding ability has laid the foundation for new opportunities in the design of novel collagen mimetic complexes. The review summarizes the synthesis and integrated biophysical analyses of the structures of these collagen mimetics. Solid phase segment condensation techniques have been utilized for the synthesis of the single chain and template-assembled analogues. The characterization of the collagen-like structures has been established by temperature-dependent optical rotation measurements. CD, NMR spectroscopy, and molecular modelling simulations.