Protective effects of L- and D-carnosine on alpha-crystallin amyloid fibril formation: implications for cataract disease.
Mildly denaturing conditions induce bovine alpha-crystallin, the major structural lens protein, to self-assemble into fibrillar structures in vitro. The natural dipeptide l-carnosine has been shown to have potential protective and therapeutic significance in many diseases. Carnosine derivatives have been proposed as potent agents for ophthalmic therapies of senile cataracts and diabetic ocular complications. Here we report the inhibitory effect induced by the peptide (l- and d-enantiomeric form) on alpha-crystallin fibrillation and the almost complete restoration of the chaperone activity lost after denaturant and/or heat stress. Scanning force microscopy (SFM), thioflavin T, and a turbidimetry assay have been used to determine the morphology of alpha-crystallin aggregates in the presence and absence of carnosine. DSC and a near-UV CD assay evidenced that the structural precursors of amyloid fibrils are polypeptide chain segments that lack stable structural elements. Moreover, we have found a disassembling effect of carnosine on alpha-crystallin amyloid fibrils. Finally, we show the ability of carnosine to restore most of the lens transparency in organ-cultured rat lenses exposed to similar denaturing conditions that were used for the in vitro experiments.
Institute of Biostructures and Bioimaging (IBB), CNR, 95125 Catania, Italy.
Francesco
F
Attanasio
Sebastiano
S
Cataldo
Salvatore
S
Fisichella
Silvia
S
Nicoletti
Vincenzo Giuseppe
VG
Nicoletti
Bruno
B
Pignataro
Anna
A
Savarino
Enrico
E
Rizzarelli
D000682:Amyloid / Q000737:chemistry*; D000818:Animals; D002152:Calorimetry, Differential Scanning; D002336:Carnosine / Q000737:chemistry* / Q000494:pharmacology; D002386:Cataract / Q000378:metabolism*; D002417:Cattle; D002942:Circular Dichroism; D005260:Female; D018625:Microscopy, Atomic Force; D018832:Molecular Chaperones / Q000378:metabolism; D009924:Organ Culture Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism; D038201:alpha-Crystallins / Q000037:antagonists & inhibitors / Q000737:chemistry*
D000682:Amyloid; D018832:Molecular Chaperones; D038201:alpha-Crystallins; D002336:Carnosine