Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2020 Jul 28;16:99. doi: 10.1186/s13007-020-00641-1

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

PMC Copyright notice

Fig. 2 — Substrate binding assay of AtGALS1. a Activity analysis of AtGASL1 using carbohydrate gel electrophoresis (PACE). Incubation of YFP-AtGALS1 microsomes and ANTS-labeled β-1,4-galactotetraose (Gal₄) in the presence and absence of 200 μM UDP-Gal displayed activity of catalyzing the elongation of galactan backbone. No activity was observed in the p19 control. b Initial fluorescence counts of YFP-AtGALS1 microsomes in MES buffer at different concentrations of UDP-Gal. The variation in fluorescence across the concentration gradients is within the tolerance range (± 10%). c Thermographs of YFP-AtGALS1 binding to UDP-Gal provide well-defined curves. The blue region at 0 s indicates cold spot before the temperature gradient was applied, and the red region at 15 s shows the hot spot during the thermophoresis. d Dose–response curve for the binding interaction between YFP-AtGALS1 and UDP-Gal by plotting Fnorm against the ligand concentration. The binding curve yields a K_d of 101 μM. e Normalized binding curve of YFP-AtGALS1 and ST-YFP control in presence of UDP-Gal. The binding curve yields a K_d of 112 ± 30 μM. Concentration of YFP-AtGALS1 or ST-YFP were kept constant while the UDP-Gal concentration varied from 2.5 mM to 76.3 nM. Data were fitted to K_d model and confidence interval of the K_d is calculated from the variance of the fitted parameter, derived from Levenberg–Marquardt fit-algorithm. Error Bar: SD, n = 3. Values are the average ± 68% confidence