Chandrika Mittal, Ram Swaroop Harsolia and Jay Kant Yadav*
Background: The cross β-sheet structures in amyloids are usually proteolytic resistant. The amyloid like protein aggregates derived from human cataracts are distinct in the sense that they do not exist as fibrils rather they are found to be aggregates of sequestered crystalline.
Methods: The proteolytic susceptibility was measured by recording changes in the Congo Red (CR) bathochromic shifts, Thioflavin T (ThT) and 8-Anilino-1-Naphthalene Sulfonic acid (ANS) fluorescence emission. The difference between the degradation patterns of these two fractions of the isolated protein was further estimated by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Furthermore, to substantiate the data regarding the proteolytic degradation of aggregated crystalline, the native PAGE has been performed which provided an evident confirmation of its degradation.
Findings: The obtained data suggest that the soluble protein is less susceptible to proteolytic cleavage than the insoluble amyloid like protein aggregates.
Conclusions: The data suggest that these aggregates are highly susceptible to proteolytic degradation; hence, it is inferred that they do not exist in a conventional amyloid like conformation.
Published Date: 2023-02-10; Received Date: 2022-09-16
