Ambient Temperature Affects Protein Self-Assembly by Interfering with the Interfacial Aggregation Behavior

Amyloid fibrillation is known to be associated with degenerativediseases, and mature fibrils are also considered as valuable biomedicalmaterials. Thus, the mechanism and influencing factors of fibrillationhave always been the focus of research. However, in vitro studiesare always plagued by low reproducibility of kinetics and the molecularmechanism of amyloid fibrillation is under debate until now. Here,we identified the ambient temperature (AT) as a non-negligible interferingfactor in in vitro self-assembly of globular protein hen egg-whitelysozyme for the first time. By multimodal molecular spectroscopymethods, not only the effect of ATs on the kinetics of protein aggregationwas described but also the conformational changes of the molecularstructure with different ATs were captured. Through investigatingthe dependence of interfacial area and catalysis, the reason for thisinfluence was construed by the various aggregation behaviors of proteinmolecules in the two-phase interface. The results suggest that invitro mechanism research on protein fibrillation needs to first clarifythe AT for a more accurate comparative analysis. The proposal of thisconcept will provide a new clue for a deeper understanding of themechanism of protein self-assembly and may have an impact on evaluatingthe efficiency of amyloid accelerators or inhibitors based on thecomparative analysis of protein self-assembly.