Quantification of protein-bound sodium dodecyl sulfate by Rhodamine B: a method for identification of kinetically stable proteins
| Autor: | Debratna Mukherjee, Reema Bhattacharya, Debasish Bhattacharyya |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Circular dichroism
Fluorophore Globular protein Biophysics Ligands Biochemistry Protein Structure Secondary chemistry.chemical_compound Rhodamine B Animals Denaturation (biochemistry) Sodium dodecyl sulfate Molecular Biology Protein secondary structure chemistry.chemical_classification Chromatography Protein Stability Rhodamines Circular Dichroism Proteins Sodium Dodecyl Sulfate Serum Albumin Bovine Cell Biology Hydrogen-Ion Concentration Reference Standards Catalase Fluorescence Kinetics Spectrometry Fluorescence chemistry |
| Zdroj: | Analytical biochemistry. 417(1) |
| ISSN: | 1096-0309 |
| Popis: | Sodium dodecyl sulfate (SDS) bound to proteins in solution could be estimated by passing through Extracti-Gel that removes free SDS followed by specific interaction of the fluorophore Rhodamine B with protein-bound SDS. The resulting fluorescence intensity is compared with a calibration curve. Whereas globular proteins respond to binding of 1.4 mg SDS/mg protein under native conditions, “kinetically stable” proteins that are otherwise resistant to denaturation due to structural integrity show a low level of SDS binding. Analysis of the circular dichroism spectrum shows that in spite of the low level of SDS binding to kinetically stable proteins under nondenaturing conditions, the detergent generates considerable secondary structure in these proteins. Because the low level of SDS binding is a general feature of kinetically stable proteins, the protocol may fulfill one of the criteria to classify a protein as kinetically stable. |
| Databáze: | OpenAIRE |
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