title: Role of the clathrin terminal domain in regulating coated pit dynamics revealed by small molecule inhibition.
abstract: Clathrin-mediated endocytosis (CME) regulates many cell physiological processes such as the internalization of growth factors and receptors, entry of pathogens, and synaptic transmission. Within the endocytic network, clathrin functions as a central organizing platform for coated pit assembly and dissociation via its terminal domain (TD). We report the design and synthesis of two compounds named pitstops that selectively block endocytic ligand association with the clathrin TD as confirmed by X-ray crystallography. Pitstop-induced inhibition of clathrin TD function acutely interferes with receptor-mediated endocytosis, entry of HIV, and synaptic vesicle recycling. Endocytosis inhibition is caused by a dramatic increase in the lifetimes of clathrin coat components, including FCHo, clathrin, and dynamin, suggesting that the clathrin TD regulates coated pit dynamics. Pitstops provide new tools to address clathrin function in cell physiology with potential applications as inhibitors of virus and pathogen entry and as modulators of cell signaling.
authors.affiliation: Institute of Chemistry and Biochemistry & Neurocure Cluster of Excellence, Freie Universität Berlin, 14195 Berlin, Germany.
authors.forename: Lisa
authors.initials: L
authors.lastname: von Kleist
authors.forename: Wiebke
authors.initials: W
authors.lastname: Stahlschmidt
authors.forename: Haydar
authors.initials: H
authors.lastname: Bulut
authors.forename: Kira
authors.initials: K
authors.lastname: Gromova
authors.forename: Dmytro
authors.initials: D
authors.lastname: Puchkov
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Robertson
authors.forename: Kylie A
authors.initials: KA
authors.lastname: MacGregor
authors.forename: Nikolay
authors.initials: N
authors.lastname: Tomilin
authors.forename: Nikolay
authors.initials: N
authors.lastname: Tomlin
authors.forename: Arndt
authors.initials: A
authors.lastname: Pechstein
authors.forename: Ngoc
authors.initials: N
authors.lastname: Chau
authors.forename: Megan
authors.initials: M
authors.lastname: Chircop
authors.forename: Jennette
authors.initials: J
authors.lastname: Sakoff
authors.forename: Jens Peter
authors.initials: JP
authors.lastname: von Kries
authors.forename: Wolfram
authors.initials: W
authors.lastname: Saenger
authors.forename: Hans-Georg
authors.initials: HG
authors.lastname: Kräusslich
authors.forename: Oleg
authors.initials: O
authors.lastname: Shupliakov
authors.forename: Phillip J
authors.initials: PJ
authors.lastname: Robinson
authors.forename: Adam
authors.initials: A
authors.lastname: McCluskey
authors.forename: Volker
authors.initials: V
authors.lastname: Haucke
mesh_terms: D033962:Adaptor Protein Complex 2 / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D002966:Clathrin / Q000737:chemistry* / Q000378:metabolism*; D003034:Coated Pits, Cell-Membrane / Q000187:drug effects / Q000378:metabolism*; D018360:Crystallography, X-Ray; D003584:Cytological Techniques / Q000379:methods*; D034281:Dynamins / Q000378:metabolism; D004705:Endocytosis; D006801:Humans; D051379:Mice; D017434:Protein Structure, Tertiary; D015398:Signal Transduction; D054852:Small Molecule Libraries; D013569:Synapses / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D033962:Adaptor Protein Complex 2; D002966:Clathrin; D054852:Small Molecule Libraries; D034281:Dynamins