Journal: eLife

Article Title: Thalamocortical axons control the cytoarchitecture of neocortical layers by area-specific supply of VGF

doi: 10.7554/eLife.67549

Figure Lengend Snippet:

Article Snippet: For electroporation-based cell ablation, pCAG- DTR was constructed as follows: a cDNA fragment containing the coding region of human HBEGF (GenBank accession number: BC033097) was obtained from a commercial supplier (clone ID 100067676, DNAFORM).

Techniques: CRISPR, Recombinant, Plasmid Preparation, Expressing, Sequencing, Imaging, Software, Staining

Journal: Journal of Lipid Research

Article Title: Characterization of homologous sphingosine-1-phosphate lyase isoforms in the bacterial pathogen Burkholderia pseudomallei [S]

doi: 10.1194/jlr.M071258

Figure Lengend Snippet: Metabolism of S1P. S1PL catalyzes the degradation of S1P to 2E-HEX and PE. The 2E-HEX product is a substrate for the NAD + -dependent enzyme, FALDH, which oxidizes 2E-HEX to the fatty acid with the concomitant reduction of the NAD + cofactor.

Article Snippet: The full-length FALDH gene sequence (coding isoform 2) was purchased as a cDNA clone from Imagenes in a pOTB7 vector (UniProt code, P51648).

Techniques:

Journal: Journal of Lipid Research

Article Title: Characterization of homologous sphingosine-1-phosphate lyase isoforms in the bacterial pathogen Burkholderia pseudomallei [S]

doi: 10.1194/jlr.M071258

Figure Lengend Snippet: Kinetic analysis of FALDH and S1PL2021/S1PL2025. A: Recombinant human FALDH (0.5 μM) displays enzyme activity with aldehyde substrates, 2E-HEX, hexadecanal, and tetradecanal. B: Michaelis-Menten kinetic analysis of FALDH using 2E-HEX as a substrate (0–260 μM). The K M for 2E-HEX = 19.4 ± 3.2 μM and k cat = 0.22 ± 0.01 s −1 . C, D: Michaelis-Menten kinetic analysis using the S1PL/FALDH coupled assays with S1P as substrate (6.0 μM S1PL, 0.5 μM FALDH). The K M and k cat values for S1P are: 48.2 ± 9.4 μM and 0.015 ± 0.003 s −1 for (C) S1PL2021 and 50.2 ± 8.3 μM and 0.008 ± 0.0005 s −1 for (D) S1PL2025. E: The S1PL2021 K271A mutant displays negligible activity when using the coupled assay. F: Use of the coupled assay to allow comparison of the S1PL20121 lyase activity with S1P (100 μM) and D- erythro sphingosine (300 μM) substrates.

Article Snippet: The full-length FALDH gene sequence (coding isoform 2) was purchased as a cDNA clone from Imagenes in a pOTB7 vector (UniProt code, P51648).

Techniques: Recombinant, Activity Assay, Mutagenesis, Comparison

Journal: Journal of Lipid Research

Article Title: Characterization of homologous sphingosine-1-phosphate lyase isoforms in the bacterial pathogen Burkholderia pseudomallei [S]

doi: 10.1194/jlr.M071258

Figure Lengend Snippet: Michaelis-Menten kinetic data for recombinant human FALDH using 2E-HEX as substrate and the B. pseudomallei S1PL2021 and S1PL2025 enzymes using S1P as a substrate

Article Snippet: The full-length FALDH gene sequence (coding isoform 2) was purchased as a cDNA clone from Imagenes in a pOTB7 vector (UniProt code, P51648).

Techniques: Recombinant