A Single-Molecule Assay to Directly Identify Solvent-Accessible Disulfide Bonds and Probe Their Effect on Protein Folding

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• 作者：Sri Rama Koti Ainavarapu ; Arun P. Wiita ; Hector H. Huang ; Julio M. Fernandez
• 刊名：Journal of the American Chemical Society
• 出版年：2008
• 出版时间：January 16, 2008
• 年：2008
• 卷：130
• 期：2
• 页码：436 - 437
• 全文大小：99K
• 年卷期：v.130,no.2(January 16, 2008)
• ISSN：1520-5126

文摘

Mechanical unfolding of a single domain protein with engineered disulfide bonds in reducing and oxidizing conditions has been probed by single-molecule force spectroscopy. Reduction of the solvent-accessible disulfide bond by the chemical reducing agent DTT resulted in an increase in the protein contour length, and the kinetics of reduction were measured at the single-molecule level. In contrast, a sequestered disulfide bond was not readily reduced by DTT. We also show that the protein folding kinetics are sensitive to the redox environment, and the differential kinetics of the oxidized and reduced domains were simultaneously measured.