Conversion of yeast phosphoglycerate kinase into amyloid-like structure

• Published in: Proteins, structure, function, and bioinformatics structure, function, and bioinformatics, 2000-05, Vol.39 (3), p.204-211
• Main Authors: Damaschun, Gregor, Damaschun, Hilde, Fabian, Heinz, Gast, Klaus, Kröber, Reinhard, Wieske, Martin, Zirwer, Dietrich
• Format: Article
• Language: English
• Subjects: amyloid; Amyloid - chemistry; Amyloid - metabolism; Amyloid - ultrastructure; Congo Red; dynamic light scattering; electron microscopy; fibrillogenesis; infrared spectroscopy; Light; Microscopy, Electron; misfolding; phosphoglycerate kinase; Phosphoglycerate Kinase - chemistry; Phosphoglycerate Kinase - metabolism; Phosphoglycerate Kinase - ultrastructure; Protein Folding; Protein Structure, Quaternary; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae - ultrastructure; Scattering, Radiation; Spectrophotometry, Infrared; X-ray scattering; X-Rays
• ISSN: 0887-3585; 1097-0134
• DOI: 10.1002/(SICI)1097-0134(20000515)39:3<204::AID-PROT20>3.0.CO;2-8

Yeast phosphoglycerate kinase is a structurally well‐characterized enzyme consisting of 415 amino acids without disulfide bonds. Anion‐induced refolding from its acid‐unfolded state gives rise to the formation of worm‐like amyloid fibrils with a persistence length of 73 nm. Electron microscopy and small‐angle X‐ray scattering data indicate that the fibrils have an elliptical cross‐section with dimensions of 10.2 nm × 5.1 nm. About half of all amino acids are organized in form of cross‐β structure which gives rise to typical infrared spectra, X‐ray diffraction and yellow‐green birefringence after Congo red staining. The kinetics of amyloid formation, monitored by infrared spectroscopy, dynamic light scattering and X‐ray scattering, was found to be strongly dependent on protein concentration. The infrared data indicate that the formation of cross‐β structure practically comes to an end already after some hours, whereas the length‐growth of the amyloid fibrils, monitored by small‐angle X‐ray scattering, was not yet completed after 1,300 hours. Proteins 2000;39:204–211. © 2000 Wiley‐Liss, Inc.