Molecular and supramolecular studies on polyglycine and poly-l-proline

Abstract

Elastin is a cross-linked protein, whose soluble precursor is tropoelastin, responsible for resilience and elastic recoil in vertebrate tissues. Glycine and proline are among the most repeated amino acids in tropoelastin primary structure, the high flexible glycine being present 222 times and the more constrained proline being present 96 times. In order to deeper investigate the role of glycine and proline residues in elastin, we studied the molecular and supramolecular structures of polyglycine and poly-L-proline homopolypeptides as significant sequences for the protein. As a matter of fact, up to now, if few conformational studies are accessible only for poly-L-proline homopolypeptide in solution and for polyglycine homopolypeptide in the solid state, limited supramolecular studies are available for both homopolypeptides. Given the self-aggregation properties of these homopolypeptides, we investigated the aggregation mechanism by turbidimetry measurements together with Congo red birefringence assay, ThT fluorescence spectroscopy, and atomic force microscopy and transmission electron microscopy studies. At molecular level, we show the dominance of the cross-β structure for polyglycine fibrils while for poly-L-proline aggregates PPII conformation prevails. At supramolecular level, the results show that polyglycine is able to self-aggregate into amyloid-like fibres while poly-L-proline aggregates by following a specific pathway ranging from protofibrils to fibrils. These findings suggest that the self-aggregation properties of elastin are influenced by tropoelastin primary structure thus explaining why glycine-rich elastin-derived polypeptide sequences are amyloidogenic (Gly-effect) while proline-rich elastin-derived polypeptide sequences (Pro-effect) are able to coacervate.