Effect of trehalose on W7FW14F apomyoglobin and insulin fibrillization: new insight into inhibition activity.
Trehalose, a disaccharide present in many nonmammalian species, protects cells against various environmental stresses. Trehalose has recently been shown to decrease aggregate formation and toxicity in cell models and to alleviate amyloid-induced diseases. The aim of our study was to use two amyloid-forming proteins, i.e., W7FW14F apomyoglobin and insulin, as model systems to elucidate the molecular mechanism by which trehalose affects the amyloid aggregation process and to investigate further its therapeutic potential. Protein aggregation was examined by far-UV circular dichroism, UV absorption, thioflavin T fluorescence, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, atomic force microscopy, and Fourier transform infrared spectroscopy. Cell viability was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay. We found that trehalose does not inhibit protein aggregation but acts at different stages of the fibrillization process depending on the protein model used. In fact, trehalose dose-dependently inhibited fibril formation in the W7FW14F apomyoglobin model and increased the lag phase in the insulin model. In both cases, trehalose caused accumulation of toxic oligomeric species. The results suggest that trehalose may favor or inhibit the formation of "on-pathway" or "off-pathway" oligomeric intermediates depending on the nature of the aggregating protein.
Vilasi
Silvia
S
Dipartimenti di Biochimica e Biofisica and Medicina Sperimentale, Seconda Università degli Studi di Napoli, Via L. De Crecchio 7, 80138 Napoli, Italy.
Iannuzzi
Clara
C
Portaccio
Marianna
M
Irace
Gaetano
G
Sirangelo
Ivana
I
D016475:3T3 Cells; D000818:Animals; D001059:Apoproteins / Q000737:chemistry / Q000187:drug effects*; D002417:Cattle; D002942:Circular Dichroism; D007328:Insulin / Q000737:chemistry*; D051379:Mice; D018625:Microscopy, Atomic Force; D009211:Myoglobin / Q000737:chemistry / Q000187:drug effects*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D017550:Spectroscopy, Fourier Transform Infrared; D014199:Trehalose / Q000494:pharmacology*
D001059:Apoproteins; D007328:Insulin; D009211:Myoglobin; C010857:apomyoglobin; D014199:Trehalose