Journal: PLoS ONE

Article Title: Spastin's Microtubule-Binding Properties and Comparison to Katanin

doi: 10.1371/journal.pone.0050161

Figure Lengend Snippet: Movie S2 shows GFP-katanin p60 (green) moving along AlexaFluor 555 microtubules (red). The movie plays in real time, the width of the canvas is 25.1 µm.

Article Snippet: A codon-optimized version of full-length human katanin was synthesized commercially (MorphoSys/Sloning; Planegg, Germany).

Techniques:

Journal: PLoS ONE

Article Title: Spastin's Microtubule-Binding Properties and Comparison to Katanin

doi: 10.1371/journal.pone.0050161

Figure Lengend Snippet: The figure shows a sequence alignment of domains of Drosophila and human spastin, as well as human katanin. The black contour highlights the position of the human spastin linker, residues with a high degree of conservation are red, residues with low degree are blue. The location of the highly basic patch in human spastin is boxed in blue. Above the alignment, the location of the sequence in the context of human spastin is indicated, below the location in human katanin. For details, see text.

Article Snippet: A codon-optimized version of full-length human katanin was synthesized commercially (MorphoSys/Sloning; Planegg, Germany).

Techniques: Sequencing

Journal: PLoS ONE

Article Title: Spastin's Microtubule-Binding Properties and Comparison to Katanin

doi: 10.1371/journal.pone.0050161

Figure Lengend Snippet: Nucleotide dependence of microtubule interaction.

Article Snippet: A codon-optimized version of full-length human katanin was synthesized commercially (MorphoSys/Sloning; Planegg, Germany).

Techniques:

Journal: PLoS ONE

Article Title: Spastin's Microtubule-Binding Properties and Comparison to Katanin

doi: 10.1371/journal.pone.0050161

Figure Lengend Snippet: Salt dependence of microtubule interaction.

Article Snippet: A codon-optimized version of full-length human katanin was synthesized commercially (MorphoSys/Sloning; Planegg, Germany).

Techniques:

Journal: PLoS ONE

Article Title: Spastin's Microtubule-Binding Properties and Comparison to Katanin

doi: 10.1371/journal.pone.0050161

Figure Lengend Snippet: Panel A displays the constructs used for katanin binding experiments. Panel B shows a quantitative SDS-gel of supernatants (unbound) and pellets (microtubule-bound) of an in vitro binding assay of truncated katanin constructs and microtubules. Increasing microtubule concentrations (0 to 10 µM) were incubated with a fixed katanin construct concentration (1 µM). The density of the katanin construct band was plotted against the microtubule concentration and fitted to a Hill function (panel C). The half-maximal saturation was reached at 0.34 µM (Kat12) and 0.40 µM (Kat2). Only constructs containing domain 2 were able to bind to microtubules.

Article Snippet: A codon-optimized version of full-length human katanin was synthesized commercially (MorphoSys/Sloning; Planegg, Germany).

Techniques: Construct, Binding Assay, SDS-Gel, In Vitro, Incubation, Concentration Assay

Journal: PLoS ONE

Article Title: Spastin's Microtubule-Binding Properties and Comparison to Katanin

doi: 10.1371/journal.pone.0050161

Figure Lengend Snippet: Binding stoichiometry.

Article Snippet: A codon-optimized version of full-length human katanin was synthesized commercially (MorphoSys/Sloning; Planegg, Germany).

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Spastin's Microtubule-Binding Properties and Comparison to Katanin

doi: 10.1371/journal.pone.0050161

Figure Lengend Snippet: Panel A shows a SDS-gel of co-sedimentation assays with katanin (E309Q mutant, 1 mM ATP) and a constant concentration of microtubules (2 µM; indicated by a dotted line). With increasing katanin concentrations, an increasing amount of protein is co-sedimented with microtubules. Panel B: Plot of the densitometric analysis as in . Panel C shows the same experiment for wild type katanin.

Article Snippet: A codon-optimized version of full-length human katanin was synthesized commercially (MorphoSys/Sloning; Planegg, Germany).

Techniques: SDS-Gel, Sedimentation, Mutagenesis, Concentration Assay