Formation of amyloid fibrils by bovine carbonic anhydrase.
Amyloids are typically characterized by extensive aggregation of proteins where the participating polypeptides are involved in formation of intermolecular cross beta-sheet structures. Alternate structure attainment and amyloid formation has been hypothesized to be a generic property of a polypeptide, the propensities of which vary widely depending on the polypeptide involved and the physicochemical conditions it encounters. Many proteins that exist in the normal form in-vivo have been shown to form amyloid when incubated in partially denaturing conditions. The protein bovine carbonic anhydrase II (BCA II) when incubated in mildly denaturing conditions showed that the partially unfolded conformers assemble together and form ordered amyloid aggregates. The properties of these aggregates were tested using the traditional Congo-Red (CR) and Thioflavin-T (ThT) assays along with fluorescence microscopy, transmission electron microscopy (TEM), and circular dichroism (CD) spectroscopy. The aggregates were found to possess most of the characteristics ascribed to amyloid fibers. Thus, we report here that the single-domain globular protein, BCA II, is capable of forming amyloid fibrils. The primary sequence of BCA II was also analyzed using recurrence quantification analysis in order to suggest the probable residues responsible for amyloid formation.
Rana
Anshul
A
Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology, Hauz Khas. New Delhi 110016, India.
Gupta
Teemish Praveen
TP
Bansal
Saurabh
S
Kundu
Bishwajit
B
D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D000818:Animals; D052160:Benzothiazoles; D024402:Carbonic Anhydrase II / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D002417:Cattle; D002942:Circular Dichroism; D003224:Congo Red / Q000737:chemistry; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D017510:Protein Folding; D013844:Thiazoles / Q000737:chemistry
D000682:Amyloid; D052160:Benzothiazoles; D013844:Thiazoles; C009462:thioflavin T; D003224:Congo Red; D024402:Carbonic Anhydrase II