Journal: Redox Biology

Article Title: Identification and quantification of sites of nitration and oxidation in the key matrix protein laminin and the structural consequences of these modifications

doi: 10.1016/j.redox.2019.101226

Figure Lengend Snippet: Nitration reduces cell adhesion to laminin-derived C16 peptide. A) Crystal structure of the N-terminal domain of the laminin- γ 1 chain shows solvent exposure of Tyr-145 in the C16 sequence (structure from Ref. ). The C16 peptide was left untreated (0X) or nitrated with tetranitromethane (TNM), either in 10-fold (10X) or 100-fold (100X) molar excess to the peptide, and used to coat 96-well cell culture plates. HCAECs stained with Calcein-AM were allowed to adhere to the pre-coated 96-well plates for 90 min. After removal of nonadherent cells, the adherent cells were quantified using Calcein-AM fluorescence. Values are expressed as means ± SEM (n = 3). Asterisks indicate significant difference from control samples (ANOVA with Dunnett's correction for multiple testing, ***p < 0.001).

Article Snippet: Synthetic C16 peptide (KAFDITYVRLKF, synthesized by EZBiolab, Carmel, IN, USA) used for the cell adhesion assay was incubated with tetranitromethane (TNM) to give the peptide nitrated at the sole Tyr residue in high yields with minimal site products.

Techniques: Nitration, Derivative Assay, Solvent, Sequencing, Cell Culture, Staining, Fluorescence, Control