Citation: ZHANG Yu-Jiao, TANG Qian, CAO Hong-Yu, ZHENG Xue-Fang. Acid-Induced Unfolding Process of Myoglobin and Its Mutant under Macromolecular Crowding Conditions[J]. Acta Physico-Chimica Sinica, ;2013, 29(08): 1785-1792. doi: 10.3866/PKU.WHXB201305271 shu

Acid-Induced Unfolding Process of Myoglobin and Its Mutant under Macromolecular Crowding Conditions

1.
1 School of Life Science and Biotechnology, Dalian University, Dalian 116622, Liaoning Province, P. R. China;
2 Liaoning Key Lab of Bio-organic Chemistry Dalian University, Dalian 116622, Liaoning Province, P. R. China

Received Date: 18 March 2013
Available Online: 27 May 2013
Fund Project: 国家自然科学基金(21271036, 20871024)资助项目 (21271036, 20871024)

In this paper, we studied the unfolding process of myoglobin (Mb) and its mutant Mb(D60K) induced by acid under macromolecular crowding conditions, using ultraviolet-visible absorption, synchronous fluorescence, and circular dichroism (CD) spectroscopies. The spectroscopic data showed that, with the addition of Dextran70 or Ficoll70, the denaturation midpoint of Mb(WT) decreased from 4.25 to 3.78 or 3.76, respectively. In addition, the acid tolerance of Mb(WT) improved under macromolecular crowding conditions. The denaturation midpoint of Mb(D60K) was 4.19, which was a slight decrease from that of Mb (4.25). Upon addition of Dextran70 or Ficoll70, the denaturation midpoint of Mb(D60K) decreased from 4.19 to 3.74 or 3.12, respectively. The spectroscopic results illustrated that the amino acid mutant and crowding agents could stabilize the microenvironment surrounding the heme and aromatic amino acid as well as the second structure of Mb, protecting its native state.
- Crowding agent,
- Acid denaturation,
- Myoglobin,
- Mutant,
- Spectrometry
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