N-Ethylmaleimide influenced the evaluation of disulfide cross-links in the oxidized myofibrillar proteins using the non-reducing SDS-PAGE
https://doi.org/10.21323/2414-438X-2022-7-2-91-96
Abstract
The present study aimed to investigate the effect of N-Ethylmaleimide (NEM) on the evaluation of disulfide formation in the oxi dized myofibrillar proteins during the sample preparation of the non-reducing SDS-PAGE procedure. For this purpose, extracted myofibrillar proteins were oxidized firstly via a Fenton oxidation reaction, and non-oxidized proteins were used as a control. Before running SDS-PAGE, in the sample preparation, these oxidized and non-oxidized proteins were prepared according to the three dif ferent sample preparation methods with or without the presence of N-Ethylmaleimide or β-mercaptoethanol. Results showed that oxidized proteins treated with NEM regardless of sample preparation methods presented attenuated bands of myosin heavy chain monomer in the non-reducing SDS-PAGE gels, suggesting that the disulfide bonds formed as a result of protein oxidation could be preserved by NEM during sample preparation. Meanwhile, a possible mechanism for the effect of NEM was proposed.
Supporting agencies: This research was funded by the National Natural Science Foundation of China, grant number 31701671 and the Dalian Science and Technology Bureau Fund, grant No. 2021RQ019
About the Authors
W. Wang
School of Food Science and Technology, Dalian Polytechnic University
China
China
Wenhui Wang, Master student
Ganjingzi #1, 116304, Dalian, Liaoning
S. Wang
School of Food Science and Technology, Dalian Polytechnic University
China
China
Shouyin Wang, Master student
Ganjingzi #1, 116304, Dalian, Liaoning
S. Li
School of Food Science and Technology, Dalian Polytechnic University;
National Engineering Research Center of Seafood
China
China
Shengjie Li, PhD, Associate Professor, School of Food Science and Technology, Dalian Polytechnic University, National Engineering Research Center of Seafood
Ganjingzi #1, 116304, Dalian, Liaoning
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Review
For citations:
Wang W., Wang S., Li S. N-Ethylmaleimide influenced the evaluation of disulfide cross-links in the oxidized myofibrillar proteins using the non-reducing SDS-PAGE. Theory and practice of meat processing. 2022;7(2):91-96. https://doi.org/10.21323/2414-438X-2022-7-2-91-96
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