Abstract
Hyperglycemic condition i.e. an increase in blood glucose concentration has been linked to bring about structural alterations in the native state of proteins. Glucose concentrations of 50 and 100 mM in vitro, which correspond to hyperglycemic condition, were tested to investigate their effect on lysozyme native structure. Incubating enzyme with 50 and 100 mM glucose for a period of 7 days, an intermediate state on day 4 and 3 was observed, respectively. The presence of intermediate state was characterized by a 22 % increase in the intrinsic fluorescence intensity with a red shift of 20 nm compared to the native state, a 5 % increase in ANS-fluorescence intensity relative to the native due to the surfacing of hydrophobic clusters and a sharp decrease in near-UV CD signal at around 284 and 291 nm. The state retains substantial native-like secondary structure. This partially unfolded intermediate state can be referred as 'molten globule', which finally tends to aggregate on day 6 and 4 with 50 and 100 mM glucose concentration, respectively, as a result of cross-linking between lysozyme molecules. The aggregates were confirmed by the presence of β-sheet structure as depicted by far-UV CD, an increase in ThT fluorescence as well as the fibrillar morphology shown by SEM. Moreover, advanced glycation end products were also accompanied as the emission peak was observed at 460 and 470 nm corresponding to the formation of pentosidine and malonaldehyde, respectively.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.