Forbidden Secondary Structures Found in Gel-Forming Fibrils of Glycylphenylalanylglycine

• Published in: The journal of physical chemistry. B 2022-10, Vol.126 (40), p.8080-8093
• Main Authors: O’Neill, Nichole, Lima, Thamires A., Ferreira, Fabio Furlan, Thursch, Lavenia, Alvarez, Nicolas, Schweitzer-Stenner, Reinhard
• Format: Article
• Language: English
• Subjects: Amides - chemistry; B: Soft Matter, Fluid Interfaces, Colloids, Polymers, and Glassy Materials; Circular Dichroism; Peptides - chemistry; Powders; Protein Structure, Secondary; Spectroscopy, Fourier Transform Infrared
• ISSN: 1520-6106; 1520-5207
• DOI: 10.1021/acs.jpcb.2c05010

The zwitterionic l-tripeptide glycylphenylalanylglycine self-assembles into very long crystalline fibrils in an aqueous solution, which causes the formation of an exceptionally strong gel phase (G′ ∼ 5 × 106 Pa). The Rietveld refinement analysis of its powder X-ray diffraction (PXRD) pattern reveals a unit cell with four peptides forming a P212121 space group and adopting an inverse polyproline II conformation, that is, a right-handed helical structure that occupies the “forbidden” region of the Ramachandran plot. This unusual structure is stabilized by a plethora of intermolecular interactions facilitated by the large number of different functional groups of the unblocked tripeptide. Comparisons of simulated and experimental Fourier transform infrared and vibrational circular dichroism (VCD) amide I′ profiles corroborate the PXRD structure. Our experimental setup reduces the sample to a quasi-two-dimensional network of fibrils. We exploited the influence of this reduced dimensionality on the amide I VCD to identify the main fibril axis. We demonstrate that PXRD, vibrational spectroscopy, and amide I simulations provide a powerful toolset for secondary structure and fibril axis determination.