title: Oligomerization state of dynamin 2 in cell membranes using TIRF and number and brightness analysis.
abstract: Dynamin 2 is an ubiquitously expressed ∼100 kDa GTPase involved in receptor-mediated endocytosis, Golgi budding, and cytoskeletal reorganization. Dynamin molecules assemble around the necks of budding vesicles and constrict membranes in a GTP-dependent process, resulting in vesicle release. The oligomerization state of dynamin 2 in the membrane is still controversial. We investigated dynamin 2 within the plasma membrane of live cells using total internal reflection microscopy coupled with number and brightness analysis. Our results demonstrate that dynamin 2 is primarily tetrameric throughout the entire cell membrane, aside from punctate structures that may correspond to regions of membrane vesiculation.
authors.affiliation: Department of Cell and Molecular Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii.
authors.forename: Justin A
authors.initials: JA
authors.lastname: Ross
authors.affiliation: Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, California.
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Digman
authors.affiliation: Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine, California.
authors.forename: Enrico
authors.initials: E
authors.lastname: Gratton
authors.affiliation: Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Albanesi
authors.affiliation: Department of Cell and Molecular Biology, John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii. Electronic address: djameson@hawaii.edu.
authors.forename: David M
authors.initials: DM
authors.lastname: Jameson
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D034285:Dynamin II / Q000737:chemistry* / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D020836:Protein Structure, Quaternary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D034285:Dynamin II