Deamidation Slows Curli Amyloid-Protein Aggregation

Biochemistry
Hanliu WangMichael L Gross

Abstract

Nonenzymatic deamidation of asparagine and glutamine in peptides and proteins is a frequent modification both in vivo and in vitro. The biological effect is not completely understood, but it is often associated with protein degradation and loss of biological function. Here we describe the deamidation of CsgA, the major protein subunit of curli, which are important proteinaceous components of biofilms. CsgA has a high content of Asn and Gln, a feature seen in a few proteins that self-aggregate. We have implemented an approach to monitor deamidation rapidly by following the globally centroid mass shift, providing guidance for studies at the residue level. From the global mass measurement, we identified, using LC-MS/MS, extensive deamidation of several Asn residues and discovered three "Asn-Gly" sites to be the hottest spots for deamidation. The fibrillization of deamidated CsgA was measured using thioflavin T (ThT) fluorescence, circular dichroism (CD), and a previously reported hydrogen-deuterium exchange (HDX) platform. Deamidated proteins exhibit a longer lag phase and lower final ThT fluorescence, strongly suggesting slower and less amyloid fibril formation. CD spectra show that extensively deamidated CsgA remains unstructure...Continue Reading

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Citations

Aug 29, 2018·Nanotechnology·Noémie-Manuelle Dorval CourchesneNeel S Joshi
Mar 15, 2019·Cells·Fabiana Miraglia, Emanuela Colla
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Jun 3, 2021·International Journal of Molecular Sciences·Natalia SzulcMalgorzata Kotulska

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