nonlinear curve fitting with the gaussian function in originlab 2022  (OriginLab corp)

Journal: eLife

Article Title: Homo-oligomerization of the human adenosine A 2A receptor is driven by the intrinsically disordered C-terminus

doi: 10.7554/eLife.66662

Figure Lengend Snippet: The SEC data is recorded every second as absorbance at 280 nm. The baseline is corrected to ensure uniform fitting and integration across the peaks. The areas under the curve, resulting from a multiple-Gaussian curve fit, express the population of each oligomeric species. The reported standard errors of integration are within a 95% confidence interval and are calculated from the variance of the fit, not experimental errors. The levels of high-molecular-weight oligomer and dimer are expressed relative to the monomeric population in arbitrary units. A representative calculation defining the oligomer levels is given in the box.

Article Snippet: SEC chromatograms were analyzed using OriginLab using the nonlinear curve fit (Gaussian) function.

Techniques: High Molecular Weight

Journal: eLife

Article Title: Homo-oligomerization of the human adenosine A 2A receptor is driven by the intrinsically disordered C-terminus

doi: 10.7554/eLife.66662

Figure Lengend Snippet: ( A ) Curve fitting using OriginLab of all A 2A R variants used in the main text of this study, listed by the order they appear. By default, each oligomeric peak is fitted with one curve using Gaussian distribution and displayed by different color shades, with the high-molecular-weight (HMW) oligomer eluted first (dark orange), followed by the dimer (lighter orange), followed by the monomer (lightest orange). However, the HMW oligomer peak in some cases cannot be fitted with one curve and thus is fitted with two curves instead. This discrepancy can be explained by variation in HMW oligomerization order among the variants. The identity of each peak is confirmed with western blotting. The value and error from the curve fitting of each peak are given in . ( B ) Data distribution of all variants used in this study in comparison to five experimental replicates of A 2A R-WT. The C-terminally truncated mutants are represented by different shades of green in increasing darkness corresponding to the increased length of the C-terminus, with the lightest shade representing the mutant with the shortest C-terminus (A316ΔC) and the darkest shade for the mutant with the longest C-terminus (P395ΔC). The levels of dimer and HMW oligomer are expressed relative to the monomeric population in arbitrary unit, with reported errors calculated from the variance of the fit, not experimental variation. There are significant variations in the dimer and HMW oligomer levels among the WT replicates, stemming from experimental errors. These variations are mitigated when the two parameters are added as the data distribution becomes more uniform. Also, the oligomerization levels of the WT replicates are consistently higher than the mutated and truncated variants. Figure 1—figure supplement 2—source data 1. Raw size-exclusion chromatography data of five experimental replicates of A 2A R-WT.

Article Snippet: SEC chromatograms were analyzed using OriginLab using the nonlinear curve fit (Gaussian) function.

Techniques: High Molecular Weight, Western Blot, Comparison, Mutagenesis, Size-exclusion Chromatography