Journal: The Journal of Biological Chemistry

Article Title: Allosteric Inhibition of the Neuropeptidase Neurolysin *

doi: 10.1074/jbc.M114.620930

Figure Lengend Snippet: Kinetic inhibition model comparisons for dynorphin A(1–8) and neurotensin hydrolysis

Article Snippet: Crystallographic data and refinement statistics Inhibition Kinetics Inhibition kinetics for compound R2 were determined using internally quenched fluorogenic peptide substrates having the amino acid sequenced of dynorphin A(1–8) and neurotensin (Peptides International).

Techniques: Inhibition

Journal: The Journal of Biological Chemistry

Article Title: Allosteric Inhibition of the Neuropeptidase Neurolysin *

doi: 10.1074/jbc.M114.620930

Figure Lengend Snippet: Inhibition kinetics. A, mixed inhibition kinetic model with kinetic parameters and fit of hydrolysis progress curves for B, fluorogenic dynorphin A(1–8) and C, fluorogenic neurotensin. Model fitting was done with the program DynaFit. Different R2 inhibitor concentrations are indicated by color (blue, no inhibitor; green, 0.030 μm; red, 0.090 μm; magenta, 0.270 μm), and different substrate concentrations are indicated by symbol shape (circle, 0.25 or 0.125 μm; square, 0.5 or 0.25 μm; diamond, 1.0 or 0.5 μm, triangle, 2.0 or 1.0 μm; for dynorphin A(1–8) or neurotensin, respectively). Every fifth point from the progress curve data are plotted. Enzyme concentrations were varied as the inhibitor concentration increased, so the curves do not follow a simple pattern of apparent velocities.

Article Snippet: Crystallographic data and refinement statistics Inhibition Kinetics Inhibition kinetics for compound R2 were determined using internally quenched fluorogenic peptide substrates having the amino acid sequenced of dynorphin A(1–8) and neurotensin (Peptides International).

Techniques: Inhibition, Concentration Assay

Journal: The Journal of Biological Chemistry

Article Title: Allosteric Inhibition of the Neuropeptidase Neurolysin *

doi: 10.1074/jbc.M114.620930

Figure Lengend Snippet: Residuals for fit of the mixed inhibition model. Residuals from least squares minimized mixed inhibition models for A, dynorphin A(1–8), and B, neurotensin are plotted. The color and symbol scheme is the same as that used in Fig. 6. For both substrates, the residuals show a random distribution without indication of systematic error in the fit of the model to the data.

Article Snippet: Crystallographic data and refinement statistics Inhibition Kinetics Inhibition kinetics for compound R2 were determined using internally quenched fluorogenic peptide substrates having the amino acid sequenced of dynorphin A(1–8) and neurotensin (Peptides International).

Techniques: Inhibition

Journal: The Journal of Biological Chemistry

Article Title: Allosteric Inhibition of the Neuropeptidase Neurolysin *

doi: 10.1074/jbc.M114.620930

Figure Lengend Snippet: Kinetic parameters for mixed inhibition model

Article Snippet: Crystallographic data and refinement statistics Inhibition Kinetics Inhibition kinetics for compound R2 were determined using internally quenched fluorogenic peptide substrates having the amino acid sequenced of dynorphin A(1–8) and neurotensin (Peptides International).

Techniques: Inhibition

Journal: The Journal of Biological Chemistry

Article Title: Allosteric Inhibition of the Neuropeptidase Neurolysin *

doi: 10.1074/jbc.M114.620930

Figure Lengend Snippet: Models of substrate binding in the neurolysin active site channel. A, dynorphin A(1–8) binding model. B, neurotensin binding model. Peptides are shown in a stick representation with green carbons, and the R2 inhibitor (placed based on the crystal structure of its complex with neurolysin) is shown with magenta carbons. Peptides were docked with the FlexPepDock server in the absence of bound inhibitor, which was then superimposed on the docked structures (see the “Experimental Procedures”). The catalytic site zinc is shown as a blue sphere.

Article Snippet: Crystallographic data and refinement statistics Inhibition Kinetics Inhibition kinetics for compound R2 were determined using internally quenched fluorogenic peptide substrates having the amino acid sequenced of dynorphin A(1–8) and neurotensin (Peptides International).

Techniques: Binding Assay

Journal: Nucleic Acids Research

Article Title: Roles for APRIN (PDS5B) in homologous recombination and in ovarian cancer prediction

doi: 10.1093/nar/gkw921

Figure Lengend Snippet: Antibodies

Article Snippet: pSMC1 pS306 , Rabbit , 4029 , Cell Signaling Technology , , , 1:100 , .

Techniques:

Journal: Scientific Reports

Article Title: The relationship between apoptosis, chromatin configuration, histone modification and competence of oocytes: A study using the mouse ovary-holding stress model

doi: 10.1038/srep28347

Figure Lengend Snippet: Relative levels of acetylation (ace), dimethylation (met) or phosphorylation (pho) in different histone sites in oocytes with various chromatin configurations after OH at different temperatures for different times * .

Article Snippet: The primary antibodies included Anti-acetyl-H3 (Lys14) (1:200 dilution, Millipore,06-911), Acetyl Histone H4K12 Polyclonal Antibody (1:1000 dilution, Epigentek, A-4029), Acetyl Histone H4K16 Polyclonal Antibody (1:50 dilution, Epigentek, A-4030), Histone H3K4 Dimethyl Polyclonal Antibody (1:200 dilution, Epigentek, A-4032), Histone H3K9 Dimethyl Polyclonal Antibody (1:100 dilution, Epigentek, A-4035), Anti-phospho-Histone H3 (Ser10) Antibody, clone RR002 (1:100 dilution, Millipore, 05-598), and rabbit polyclonal anti-Fas antibody (dilution 1:100 for oocytes and 1:75 for cumulus cells, Abcam, ab82419).

Techniques:

Journal: Scientific Reports

Article Title: The relationship between apoptosis, chromatin configuration, histone modification and competence of oocytes: A study using the mouse ovary-holding stress model

doi: 10.1038/srep28347

Figure Lengend Snippet: The micrographs are laser confocal images of SN oocytes in which Hoechst 33342 (upper row) and H4K12 (lower row) were pseudo-colored blue and red, respectively. Original magnification × 200. Scale bar is 10 μm. Images in the same column are from the same oocyte observed at different optical wavelengths. The SN oocytes shown in different columns include oocytes before OH (0 h) or after OH at 39 °C for 1 h (39C1h) from wild-type (W) or gld (G) mice. The two tables show % annexin-positive oocytes and relative levels of H4K12 acetylation among different oocytes, respectively. Each treatment was repeated 4 times with each replicate containing about 30 oocytes. a–c: Values in the same column without a common letter in their superscripts differ (P < 0.05).

Article Snippet: The primary antibodies included Anti-acetyl-H3 (Lys14) (1:200 dilution, Millipore,06-911), Acetyl Histone H4K12 Polyclonal Antibody (1:1000 dilution, Epigentek, A-4029), Acetyl Histone H4K16 Polyclonal Antibody (1:50 dilution, Epigentek, A-4030), Histone H3K4 Dimethyl Polyclonal Antibody (1:200 dilution, Epigentek, A-4032), Histone H3K9 Dimethyl Polyclonal Antibody (1:100 dilution, Epigentek, A-4035), Anti-phospho-Histone H3 (Ser10) Antibody, clone RR002 (1:100 dilution, Millipore, 05-598), and rabbit polyclonal anti-Fas antibody (dilution 1:100 for oocytes and 1:75 for cumulus cells, Abcam, ab82419).

Techniques: