期刊
PROTEIN SCIENCE
卷 31, 期 2, 页码 396-406出版社
WILEY
DOI: 10.1002/pro.4231
关键词
amide proton exchange; desiccation; nuclear magnetic resonance; tardigrades
资金
- National Institutes of Health [R01GM127291, T32GM008570]
The study found that intrinsically disordered proteins (IDPs) provide better protection against dehydration-induced unfolding, mainly through electrostatic interactions formed by charge patterning and expanded conformations to protect the client proteins. These findings will help deepen our understanding of dehydration protection mechanisms and streamline the production of dehydrated proteins for expanded use in the medical, biotechnology, and chemical industries.
Extremotolerant organisms from all domains of life produce protective intrinsically disordered proteins (IDPs) in response to desiccation stress. In vitro, many of these IDPs protect enzymes from dehydration stress better than U.S. Food and Drug Administration-approved excipients. However, as with most excipients, their protective mechanism is poorly understood. Here, we apply thermogravimetric analysis, differential scanning calorimetry, and liquid-observed vapor exchange (LOVE) NMR to study the protection of two model globular proteins (the B1 domain of staphylococcal protein G [GB1] and chymotrypsin inhibitor 2 [CI2]) by two desiccation-tolerance proteins (CAHS D from tardigrades and PvLEA4 from an anhydrobiotic midge), as well as by disordered and globular protein controls. We find that all protein samples retain similar amounts of water and possess similar glass transition temperatures, suggesting that neither enhanced water retention nor vitrification is responsible for protection. LOVE NMR reveals that IDPs protect against dehydration-induced unfolding better than the globular protein control, generally protect the same regions of GB1 and CI2, and protect GB1 better than CI2. These observations suggest that electrostatic interactions, charge patterning, and expanded conformations are key to protection. Further application of LOVE NMR to additional client proteins and protectants will deepen our understanding of dehydration protection, enabling the streamlined production of dehydrated proteins for expanded use in the medical, biotechnology, and chemical industries.
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