Journal: Molecular cell

Article Title: In vivo evidence for ATPase-dependent DNA translocation by the Bacillus subtilis SMC condensin complex

doi: 10.1016/j.molcel.2018.07.006

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: NADH , Acros Organic , Cat # 271100010.

Techniques: Affinity Purification, Recombinant, Bradford Assay, Activity Assay, Software

Journal: Journal of Biological Chemistry

Article Title: A Legionella effector ADP-ribosyltransferase inactivates glutamate dehydrogenase

doi: 10.1016/j.jbc.2021.100301

Figure Lengend Snippet: Figure 2. Lart1 ADP-ribosylates fungal and protozoan isoforms of glutamate dehydrogenase. A, incorporation of [32P] from [32P]-adenylate NAD+ into an unknown 120 kDa protein in a yeast lysate by Lart1, but not the E137A mutant. B, dendrogram depicting the four classes of glutamate dehydrogenase isoforms labeled by their subunit molecular weight, adapted from Miñambres et al. (26). Each class is annotated with its phylogenetic distribution (black), monomer composition (blue), and cofactor preference (magenta). C, incorporation of [32P] from [32P]-NAD+ by WT Lart1 or the inactive E137A mutant with a panel of glutamate dehydrogenases as substrates: the two Legionella glutamate dehydrogenases (Lpg1581 and Lpg0245), human Glud2 (HsGlud2), yeast (Gdh1p and Gdh2p), and Dictyostelium Glud2 (DdGlud2). Reaction products were separated by SDS-PAGE and incorporated radioactivity visualized by autoradiography. Lpg, Legionella pneumophila gene; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis.

Article Snippet: [32P]-NAD (NEG023X250UC) was from Perkin Elmer.

Techniques: Mutagenesis, Labeling, Molecular Weight, SDS Page, Radioactivity, Autoradiography, Polyacrylamide Gel Electrophoresis

Journal: Journal of Biological Chemistry

Article Title: A Legionella effector ADP-ribosyltransferase inactivates glutamate dehydrogenase

doi: 10.1016/j.jbc.2021.100301

Figure Lengend Snippet: Figure 4. Lart1 targets a conserved arginine residue in the NAD+-binding pocket of glutamate dehydrogenase. A, fragmentation pattern of Gdh2p peptide containing phosphoribosyl-Arg800 identified by LC-MS/MS. The precursor [M+3H]3+ ion, m/z 657.30, is labeled with an asterisk (*) and was sub- jected to HCD fragmentation to generate the spectrum shown. The modification site was localized to arginine 800 highlighted in red. The b10 fragment ion containing the modified residue shows neutral loss of the phosphoribosyl group (−212 Da). B, endpoint assays depicting incorporation of [32P]-NAD+ by WT Lart1 into the indicated alanine mutants of Gdh2p. Mutation of the ADPR acceptor site R800 abolishes ADP-ribosylation. C, model of DdGlud2 built by Phyre (64) using Pyrococcus furiosus GDH (PDB 1HRD) as a template, indicating the position of the ADP-ribose acceptor residue Arg 763 (rendered in sticks). Bound NADH and glutamate (Glu), (modeled by alignment to liganded bovine GDH, PDB 6DHQ) are rendered in sticks. D, sequence logos depicting the con- servation of R800 and the surrounding residues in GDH enzymes from fungi (top, based on MAFFT alignment of 125 sequences), amoeba (middle, based on MAFFT alignment of 19 sequences), and metazoans (bottom, based on MAFFT alignment of 145 sequences). Red arrows indicate the position of the arginine targeted by Lart1. LC-MS/MS, liquid chromatography/mass spectrometry.

Article Snippet: [32P]-NAD (NEG023X250UC) was from Perkin Elmer.

Techniques: Residue, Binding Assay, Liquid Chromatography with Mass Spectroscopy, Labeling, Mutagenesis, Sequencing, Liquid Chromatography, Mass Spectrometry