Journal: Free radical biology & medicine

Article Title: Myeloperoxidase-mediated protein lysine oxidation generates 2-aminoadipic acid and lysine nitrile in vivo.

doi: 10.1016/j.freeradbiomed.2017.01.006

Figure Lengend Snippet: Fig. 7. Protein bound 2-AAA and LysCN formation requires protein lysyl residue exposure to the complete MPO/H2O2/Cl- system. HSA (1 mg/mL, 15 μM) was exposed to the complete MPO/H2O2/Cl- system, or to the indicated components of the MPO system. The production of 2-AAA LysCN and oxidation precursor lysine were quantified by LC/MS/MS as described in Materials and methods. Only under the complete MPO/H2O2/Cl- system, 2-AAA and LysCN were produced. Data values represent the mean ± S.D. of triplicate samples for an experiment performed at least 3 separate times.

Article Snippet: Nα-Boc-L-Lysine and Nα-acetyl-L-Lysine used as surrogates for protein lysyl residues were purchased from Chem-Impex International, Inc. (Wood Dale, Illinois).

Techniques: Residue, Liquid Chromatography with Mass Spectroscopy, Produced

Journal: Free radical biology & medicine

Article Title: Myeloperoxidase-mediated protein lysine oxidation generates 2-aminoadipic acid and lysine nitrile in vivo.

doi: 10.1016/j.freeradbiomed.2017.01.006

Figure Lengend Snippet: Fig. 8. Activated human neutrophils employ the MPO/H2O2/Cl- system to oxidize protein bound lysyl residues to 2-AAA and LysCN. The complete MPO system ( C) consisted of freshly harvested human neutrophils (PMN) (1×106 cells/mL) activated with 200 nM phorbol myristate acetate (PMA) in the presence of HSA (1 mg/mL). Where indicated, either PMN or PMA was omitted; or catalase (20 μg/mL) or methionine (2 mM) was included with the complete MPO system. Results are the mean ± S.D. of triplicate determinations for an experiment performed at least 3 separate times.

Article Snippet: Nα-Boc-L-Lysine and Nα-acetyl-L-Lysine used as surrogates for protein lysyl residues were purchased from Chem-Impex International, Inc. (Wood Dale, Illinois).

Techniques:

Journal: Nucleic Acids Research

Article Title: DNA and RNA editing without sequence limitation using the flap endonuclease 1 guided by hairpin DNA probes

doi: 10.1093/nar/gkaa843

Figure Lengend Snippet: FEN1 mediates hpDNA-guided specific mRNA knockdown in human cells. ( A ) Schematic diagram of hpDNA-guided knockdown in cells heterologously expressing A. fulgidus FEN1. ( B ) The construction of A. fulgidus FEN1 with NES, and the WB and IF images illustrating the successful expression of the merged protein. ( C ) The normalized fluorescence of groups transfected with FEN1-NES plus hpDNA (Hp-EGFP) targeting for EGFP, n = 3. *** P value <0.001. * P value <0.05. ( D ) The location of hpDNAs for the gene AFP and CDK9 with WB results of the control and experimental groups. n = 3. ( E ) HpSGN-mediated mRNA knockdown at predicated off-target loci. n = 2.

Article Snippet: Primary antibodies included antibodies against β-Actin (Abclonal Technology, AC026, 1:150 000), HA (Santa Cruz Biotech, sc-7392, 1:200), CDK9 (Cell Signaling Technology, C12F7, 1:2000) and AFP (Abclonal Technology, A11865, 1:1000).

Techniques: Expressing, Fluorescence, Transfection