Robertson MJ1, Deane FM1, Stahlschmidt W2, von Kleist L2, Haucke V2, Robinson PJ3, McCluskey A4.
Nat Protoc. 2014 Jul;9(7):1592-606. doi: 10.1038/nprot.2014.106. Epub 2014 Jun 12.This protocol describes the synthesis of two classes of clathrin inhibitors, Pitstop 1 and Pitstop 2, along with two inactive analogs that can be used as negative controls (Pitstop inactive controls, Pitnot-2 and Pitnot-2-100). Pitstop-induced inhibition of clathrin TD function acutely interferes with clathrin-mediated endocytosis (CME), synaptic vesicle recycling and cellular entry of HIV, whereas clathrin-independent internalization pathways and secretory traffic proceed unperturbed; these reagents can, therefore, be used to investigate clathrin function, and they have potential pharmacological applications. Read More