title: Pyrimidyn compounds: dual-action small molecule pyrimidine-based dynamin inhibitors.
abstract: Dynamin is required for clathrin-mediated endocytosis (CME). Its GTPase activity is stimulated by phospholipid binding to its PH domain, which induces helical oligomerization. We have designed a series of novel pyrimidine-based "Pyrimidyn" compounds that inhibit the lipid-stimulated GTPase activity of full length dynamin I and II with similar potency. The most potent analogue, Pyrimidyn 7, has an IC50 of 1.1 μM for dynamin I and 1.8 μM for dynamin II, making it among the most potent dynamin inhibitors identified to date. We investigated the mechanism of action of the Pyrimidyn compounds in detail by examining the kinetics of Pyrimidyn 7 inhibition of dynamin. The compound competitively inhibits both GTP and phospholipid interactions with dynamin I. While both mechanisms of action have been previously observed separately, this is the first inhibitor series to incorporate both and thereby to target two distinct domains of dynamin. Pyrimidyn 6 and 7 reversibly inhibit CME of both transferrin and EGF in a number of non-neuronal cell lines as well as inhibiting synaptic vesicle endocytosis (SVE) in nerve terminals. Therefore, Pyrimidyn compounds block endocytosis by directly competing with GTP and lipid binding to dynamin, limiting both the recruitment of dynamin to membranes and its activation. This dual mode of action provides an important new tool for molecular dissection of dynamin's role in endocytosis.
authors: Cell Signalling Unit, Children's Medical Research Institute, The University of Sydney , Sydney, NSW 2145, Australia.
authors: Andrew B
authors: AB
authors: McGeachie
authors: Luke R
authors: LR
authors: Odell
authors: Annie
authors: A
authors: Quan
authors: James A
authors: JA
authors: Daniel
authors: Ngoc
authors: N
authors: Chau
authors: Timothy A
authors: TA
authors: Hill
authors: Nick N
authors: NN
authors: Gorgani
authors: Damien J
authors: DJ
authors: Keating
authors: Michael A
authors: MA
authors: Cousin
authors: Ellen M
authors: EM
authors: van Dam
authors: Anna
authors: A
authors: Mariana
authors: Ainslie
authors: A
authors: Whiting
authors: Swetha
authors: S
authors: Perera
authors: Aimee
authors: A
authors: Novelle
authors: Kelly A
authors: KA
authors: Young
authors: Fiona M
authors: FM
authors: Deane
authors: Jayne
authors: J
authors: Gilbert
authors: Jennette A
authors: JA
authors: Sakoff
authors: Megan
authors: M
authors: Chircop
authors: Adam
authors: A
authors: McCluskey
authors: Phillip J
authors: PJ
authors: Robinson
mesh_terms: D000818:Animals; D001681:Biological Assay; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D015195:Drug Design; D034281:Dynamins / Q000037:antagonists & inhibitors*; D004705:Endocytosis / Q000187:drug effects; D005434:Flow Cytometry; D015394:Molecular Structure; D011485:Protein Binding / Q000187:drug effects; D011743:Pyrimidines / Q000138:chemical synthesis* / Q000737:chemistry* / Q000494:pharmacology; D054852:Small Molecule Libraries / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology
chemical_list: C000592896:N2-(2-(dimethylamino)ethyl)-N4-dodecyl-6-methylpyrimidine-2,4-diamine; D011743:Pyrimidines; D054852:Small Molecule Libraries; D034281:Dynamins; C030986:pyrimidine