To efficiently transport proteins into and across cellular membranes, specialized transport machineries engage in recognition events with different domains of their client proteins, forming sequential intermediate complexes. The short-lived nature of these interactions poses a big challenge in the identification of the key factors involved in transport reactions and their mechanism of action. Site-directed photocrosslinking is a powerful method for the detection and accurate mapping of interacting protein domains. This chapter describes a protocol that combines site-directed photocrosslinking to metabolic labeling of proteins and lipids as a method to characterize, with temporal and spatial resolution, the interactions of a secretory protein as it transverses the bacterial envelope.
CITATION STYLE
Ieva, R. (2017). Site-directed and time-resolved photocrosslinking in cells metabolically labeled with radioisotopes. In Methods in Molecular Biology (Vol. 1615, pp. 233–245). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7033-9_19
Mendeley helps you to discover research relevant for your work.