Parkinson’s disease (PD) is the second most prevalent age-related neurodegenerative disorder. The pathogenesis of PD, and other neurodegenerative diseases, has been inextricably linked with the amyloid fibrillar aggregation and deposition of α-synuclein. The cell has a range of defense mechanisms in place to prevent aggregation and maintain protein homeostasis (proteostasis). An important element of this proteostasis network are the molecular chaperone proteins. However, the persistence of diseases associated with α-synuclein aggregation indicates that their protective capacity can be ‘overwhelmed’ in the context of these diseases. Our work seeks to investigate the role of the small heat shock molecular chaperone proteins (sHsps) in protecting against α-synuclein aggregation. Specifically, we have examined interactions between α-synuclein and sHsps at various stages along α-synuclein’s aggregation pathway using a range of bulk and single molecule techniques. Our results demonstrate that sHsps interact transiently with aggregation-prone monomeric α-synuclein to prevent its aggregation in vitro. However, the efficiency by which sHsps prevent α-synuclein aggregation is highly dependent on the rate at which it aggregates. In addition, we have characterized the ability of the sHsps to interact with mature fibrillar aggregates formed by α-synuclein and established a physiologically relevant role for this interaction in preventing the cytotoxicity of the aggregates. By pursuing the mechanistic details of the manner by which sHsps interact with α-synuclein, we aim to uncover potential mechanism(s) by which sHsp chaperone activity may be targeted to attenuate diseases associated with α-synuclein aggregation.