Spectroscopic Characterization of Structure-Function Relationships in the Intrinsically Disordered Protein Complexin

Complexins play a critical role in the regulation of neurotransmission by regulating SNARE-mediated exocytosis of synaptic vesicles. Complexins can exert either a facilitatory or an inhibitory effect on neurotransmitter release, depending on the context, and different complexin domains contribute differently to these opposing roles. Structural characterization of the central helix domain of complexin bound to the assembled SNARE bundle provided key insights into the functional mechanism of this domain of complexin, which is critical for both complexin activities, but many questions remain, particularly regarding the roles and mechanisms of other complexin domains. Recent progress has clarified the structural properties of these additional domains, and has led to various proposals regarding how they contribute to complexin function. This chapter describes spectroscopic approaches used in our laboratory and others, primarily involving circular dichroism and solution-state NMR spectroscopy, to characterize structure within complexins when isolated or when bound to interaction partners. The ability to characterize complexin structure enables structure/function studies employing in vitro or in vivo assays of complexin function. More generally, these types of approaches can be used to study the binding of other intrinsically disordered proteins or protein regions to membrane surfaces or for that matter to other large physiological binding partners.