Study of RIP1/RIP3 Peptides Aggregation In Vitro

Abstract

This master's thesis study investigates RIP1/RIP3 peptide aggregation In Vitro. Protein aggregates play a major role in diseases and in normal physiological function. Although several studies have been done on the disease-causing protein and peptide aggregates, not much have been investigated on protein aggregates that play a role in normal physiological processes. Examples of protein aggregation in normal cellular function are the receptor interacting protein kinases 1 and 3, as well as RIP1/RIP3 aggregates complexes in cellular necroptosis. RIP1/RIP3 proteins form a protein aggregate complex during necroptosis. Although there are some studies on full kinase proteins aggregation, little has been done to investigate the potential of small peptide sequences of RIP1/RIP3 proteins on their aggregation mechanism, physicochemical properties, cellular functions, and potential biomedical applications. In this thesis, we study peptides consisting of four and twelve amino acid sequences of RIP1/ RIP3 proteins that are known to drive the betasheet formation and play a major role in the aggregation. The study was aided by biochemical, biophysical, mechanical, microscopy characterization, and cellular assays. The thesis study revealed that the 12 amino acid peptides exhibited amyloid like aggregates properties, and also showed mechano sensitive properties. Further, the aggregates induced cellular necrosis compared to apoptosis. The thesis study would aid in further studies on RIP1/RIP3 peptide and protein aggregation, which could be useful in understanding their role in multiple cellular functions and diseases.