Investigation of thermoreversible polymer networks by temperature dependent size exclusion chromatography

2017 ◽  
Vol 8 (43) ◽  
pp. 6598-6605 ◽  
Author(s):  
Josef Brandt ◽  
Johannes Lenz ◽  
Kai Pahnke ◽  
Friedrich Georg Schmidt ◽  
Christopher Barner-Kowollik ◽  
...  
Keyword(s):  
Size Exclusion Chromatography ◽  
Polymer Networks ◽  
Diels Alder ◽  
Size Exclusion ◽  
Temperature Dependent ◽  
Novel Approach ◽  
Exclusion Chromatography

We introduce a novel approach for studying thermoreversible Diels–Alder networks by Temperature Dependent SEC.

Temperature-dependent size exclusion chromatography for the in situ investigation of dynamic bonding/debonding reactions

2013 ◽  
Vol 405 (28) ◽  
pp. 8981-8993 ◽  
Author(s):  
Josef Brandt ◽  
Nathalie K. Guimard ◽  
Christopher Barner-Kowollik ◽  
Friedrich G. Schmidt ◽  
Albena Lederer
Keyword(s):  
Size Exclusion Chromatography ◽  
Size Exclusion ◽  
Temperature Dependent ◽  
In Situ Investigation ◽  
Exclusion Chromatography

scholarly journals aeBlue Chromoprotein Color is Temperature Dependent

2019 ◽  
Vol 27 (1) ◽  
pp. 74-84
Author(s):  
Jessica Tamayo-Nuñez ◽  
Javier de la Mora ◽  
Felipe Padilla-Vaca ◽  
Naurú Idalia Vargas-Maya ◽  
Ángeles Rangel-Serrano ◽  
...  
Keyword(s):  
Circular Dichroism ◽  
Size Exclusion Chromatography ◽  
Fluorescent Proteins ◽  
Size Exclusion ◽  
Temperature Dependent ◽  
Physicochemical Conditions ◽  
Color Shift ◽  
Exclusion Chromatography ◽  
Circular Dichroism Analysis ◽  
Circular Dichroïsm

Background: Marine sessile organisms display a color palette that is the result of the expression of fluorescent and non-fluorescent proteins. Fluorescent proteins have uncovered transcriptional regulation, subcellular localization of proteins, and the fate of cells during development. Chromoproteins have received less attention until recent years as bioreporters. Here, we studied the properties of aeBlue, a a 25.91 kDa protein from the anemone Actinia equina. Objective: To assess the properties of aeBlue chromoprotein under different physicochemical conditions. Method: In this article, during the purification of aeBlue we uncovered that it suffered a color shift when frozen. We studied the color shift by different temperature incubation and physicochemical conditions and light spectroscopy. To assess the possible structural changes in the protein, circular dichroism analysis, size exclusion chromatography and native PAGE was performed. Results: We uncover that aeBlue chromoprotein, when expressed from a synthetic construct in Escherichia coli, showed a temperature dependent color shift. Protein purified at 4 °C by metal affinity chromatography exhibited a pinkish color and shifts back at higher temperatures to its intense blue color. Circular dichroism analysis revealed that the structure in the pink form of the protein has reduced secondary structure at 4 °C, but at 35 °C and higher, the structure shifts to a native conformation and Far UV- vis CD spectra revealed the shift in an aromatic residue of the chromophore. Also, the chromophore retains its properties in a wide range of conditions (pH, denaturants, reducing and oxidants agents). Quaternary structure is also maintained as a tetrameric conformation as shown by native gel and size exclusion chromatography. Conclusion: Our results suggest that the chromophore position in aeBlue is shifted from its native position rendering the pink color and the process to return it to its native blue conformation is temperature dependent.

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