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Structural changes of ultrasonicated bovine serum albumin revealed by hydrogen–deuterium exchange and mass spectrometry

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Abstract

The structural changes of bovine serum albumin (BSA) under high-intensity ultrasonication were investigated by fluorescence spectroscopy and mass spectrometry. Evidence for the ultrasonication-induced conformational changes of BSA was provided by the intensity changes and maximum-wavelength shift in fluorescence spectrometry. Matrix-assisted laser desorption–ionization time-of-flight mass spectroscopy (MALDI-TOF MS) revealed the increased intensity of the peak at the charge state +5 and a newly emerged peak at charge state +6, indicating that the protein became unfolded after ultrasonication. Prevalent unfolding of BSA after ultrasonication was revealed by hydrogen–deuterium exchange coupled with mass spectrometry (HDX-MS). Increased intensity and duration of ultrasonication further promoted the unfolding of the protein. The unfolding induced by ultrasonication goes through an intermediate state similar to that induced by a low concentration of denaturant.

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Acknowledgments

This study was supported by National High Technology Research and Development Program of China (863 Program, No. 2013AA102205), National Program on Key Basic Research Project (No.2012CB126314), and Key Project for Science and Technology Innovation of Jiangxi Province (20124ACB00600).

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi, 330047, China

    Qiuting Zhang, Zongcai Tu, Hui Wang & Xiaomei Sha

  2. College of Life Science, Jiangxi Normal University, Nanchang, Jiangxi, 330022, China

    Zongcai Tu & Xiaoqin Huang

  3. Engineering Research Center for Biomass Conversion, Ministry of Education, Nanchang University, Nanchang, Jiangxi, 330047, China

    Hui Wang

  4. Department of Pathology, Albert Einstein College of Medicine, Yeshiva University, Bronx, New York, NY, 10461, USA

    Hui Xiao

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  1. Qiuting Zhang
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  4. Xiaoqin Huang
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Correspondence to Zongcai Tu or Hui Xiao.

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Zhang, Q., Tu, Z., Wang, H. et al. Structural changes of ultrasonicated bovine serum albumin revealed by hydrogen–deuterium exchange and mass spectrometry. Anal Bioanal Chem 406, 7243–7251 (2014). https://doi.org/10.1007/s00216-014-8136-6

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  • DOI: https://doi.org/10.1007/s00216-014-8136-6

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