Rifampicin does not prevent amyloid fibril formation by human islet amyloid polypeptide but does inhibit fibril thioflavin-T interactions: implications for mechanistic studies of beta-cell death.
Amyloid formation has been implicated in more than 20 different human diseases, including Alzheimer's disease, Parkinson's disease, and type 2 diabetes. The development of inhibitors of amyloid is a topic of considerable interest, both because of their potential therapeutic applications and because they are useful mechanistic probes. Recent studies have highlighted the potential use of rifampicin as an inhibitor of amyloid formation by a variety of polypeptides; however, there are conflicting reports on its ability to inhibit amyloid formation by islet amyloid polypeptide (IAPP). IAPP is the cause of islet amyloid in type 2 diabetes. We show that rifampicin does not prevent amyloid formation by IAPP and does not disaggregate preformed IAPP amyloid fibrils;, instead, it interferes with standard fluorescence-based assays of amyloid formation. Rifampicin is unstable in aqueous solution and is readily oxidized. However, the effects of oxidized and reduced rifampicin are similar, in that neither prevents amyloid formation by IAPP. Furthermore, use of a novel p-cyanoPhe analogue of IAPP shows that rifampicin does not significantly affect the kinetics of IAPP amyloid formation. The implications for the development of amyloid inhibitors are discussed as are the implications for studies of the toxicity of islet amyloid. The work also demonstrates the utility of p-cyanoPhe IAPP for the screening of inhibitors. The data indicate that rifampicin cannot be used to test the relative toxicity of IAPP fibrils and prefibril aggregates of IAPP.
Meng
Fanling
F
Department of Chemistry, State University of New York at Stony Brook, Stony Brook, New York 11794-3400, USA.
Marek
Peter
P
Potter
Kathryn J
KJ
Verchere
C Bruce
CB
Raleigh
Daniel P
DP
D000682:Amyloid / Q000037:antagonists & inhibitors / Q000633:toxicity*; D000975:Antioxidants / Q000378:metabolism; D052160:Benzothiazoles; D016923:Cell Death; D006801:Humans; D058228:Islet Amyloid Polypeptide; D007515:Islets of Langerhans / Q000166:cytology*; D046529:Microscopy, Electron, Transmission; D012293:Rifampin / Q000494:pharmacology*; D013844:Thiazoles / Q000737:chemistry* / Q000378:metabolism
D000682:Amyloid; D000975:Antioxidants; D052160:Benzothiazoles; D058228:Islet Amyloid Polypeptide; D013844:Thiazoles; C009462:thioflavin T; D012293:Rifampin