title: The assembly of Ni2+-actin: some peculiarities.
abstract: Nickel alters the organisation of highly dynamic cytoskeletal elements. In cultured cells Ni2+ causes microtubule aggregation and bundling as well as microfilament aggregation and redistribution. Here, we have analysed the effect(s) of Ni2+ on in vitro actin polymerisation. Using limited proteolysis by trypsin we have suggested that the regions around Arg-62 and Lys-68 change their conformation following Ni2+ binding to the single high-affinity site for divalent cations in the G-actin molecule. We have found that Ni2+ shortens the lag phase of actin polymerisation and increases the rate of assembly mainly because of an increased elongation rate. Ni2+ has no significant effect on the final plateau of actin polymerisation nor on the actin critical concentration. Electron microscopy revealed that actin filaments polymerised by 2 mM Ni2+ showed some tendency to lateral aggregation, being frequently formed by the cohesion of two or three filaments. Furthermore, they often appeared shorter than those of control as also confirmed by the larger amount of free filament ends as well as the faster depolymerisation rate than control.
authors.affiliation: University of Milan, Department of Biology, Laboratory of Biochemistry and Biophysics of Cytoskeleton Structures, via Celoria 26, 20133 Milan, Italy.
authors.forename: I
authors.initials: I
authors.lastname: DalleDonne
authors.forename: A
authors.initials: A
authors.lastname: Milzani
authors.forename: C
authors.initials: C
authors.lastname: Ciapparelli
authors.forename: M
authors.initials: M
authors.lastname: Comazzi
authors.forename: M R
authors.initials: MR
authors.lastname: Gioria
authors.forename: R
authors.initials: R
authors.lastname: Colombo
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000648:ultrastructure; D002499:Centrifugation, Density Gradient; D008278:Magnesium Sulfate; D008854:Microscopy, Electron; D009532:Nickel / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry; D011487:Protein Conformation; D014357:Trypsin
chemical_list: D000199:Actins; D011108:Polymers; C029938:nickel sulfate; D008278:Magnesium Sulfate; D009532:Nickel; D014357:Trypsin