Recovery of functional enzyme from amyloid fibrils.
Amyloid deposits, which accumulate in numerous diseases, are the final stage of multi-step protein conformational-conversion and oligomerization processes. The underlying molecular mechanisms are not fully understood, and particularly little is known about the reverse reaction. Here we show that phosphoglycerate kinase amyloid fibrils can be converted back into native protein. We achieved recovery with 60% efficiency, which is comparable to the success rate of the unfolding-refolding studies, and the recovered enzyme was folded, stable and fully active. The key intermediate stages in the recovery process are fibril disassembly and unfolding followed by spontaneous protein folding.
Department of Biophysics and Radiation Biology, Semmelweis University, Budapest, Hungary.
Gergely
G
Agócs
Katalin
K
Solymosi
Andrea
A
Varga
Károly
K
Módos
Miklós
M
Kellermayer
Péter
P
Závodszky
Judit
J
Fidy
Szabolcs
S
Osváth
D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism; D019411:Clinical Laboratory Techniques; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D010735:Phosphoglycerate Kinase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology*; D011489:Protein Denaturation; D017510:Protein Folding; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism; D013696:Temperature
D000682:Amyloid; D029701:Saccharomyces cerevisiae Proteins; D010735:Phosphoglycerate Kinase