Abstract
A simple and convenient method for water-insoluble protein surface hydrophobicity determination was developed and validated. The method is based on the non-covalent binding of Coomassie Brilliant Blue G-250 (CBBG) to aromatic and basic amino acid residues on the surface of proteins, generating insoluble protein-CBBG complexes that could be precipitated by centrifugation and cause a reduction in the absorbance at 585 nm in the supernatant. The amount of protein-bound CBBG is applied as an indicator of protein surface hydrophobicity. Thermally treated, water-insoluble myofibrillar proteins (MPs) were chosen to validate the proposed method. The amount of protein-bound CBBG increased in thermally treated protein samples and the results were compared with the surface hydrophobicity (S0) calculated by the commonly accepted fluorescence method using a 1-anilino-8-naphthalenesulfonate (ANS) probe and intrinsic tryptophan fluorescence. Results from the CBBG-binding method linearly correlated with the S0 of the ANS fluorescence method (R = 0.95), the maximum emission wavelength of protein intrinsic fluorescence (R = 0.96) and the intrinsic fluorescence intensity at 337 nm (R = -0.73), strongly suggesting the reliability of the proposed CBBG-binding method.
Original language | English |
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Pages (from-to) | 790-795 |
Number of pages | 6 |
Journal | Analytical Methods |
Volume | 8 |
Issue number | 4 |
DOIs | |
State | Published - Jan 28 2016 |
Bibliographical note
Publisher Copyright:© The Royal Society of Chemistry 2016.
Funding
This is publication No. 15-07-132 of the Kentucky Agricultural Experiment Station and is published with the approval of the Director. This work is supported by the National Institute of Food and Agriculture, U.S. Department of Agriculture, Hatch project 1005724. Author Cao would like to acknowledge the financial support from the China Scholarship Council (CSC) to grant him the opportunity to perform the study at the University of Kentucky.
Funders | Funder number |
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U.S. Department of Agriculture | 1005724 |
National Institute of Food and Agriculture | |
University of Kentucky | |
Kentucky Agricultural Experiment Station | |
China Scholarship Council |
ASJC Scopus subject areas
- Analytical Chemistry
- General Chemical Engineering
- General Engineering