A quantitative, high-throughput screen for protein stability.
In proteomic research, it is often necessary to screen a large number of polypeptides for the presence of stable structure. Described here is a technique (referred to as SUPREX, stability of unpurified proteins from rates of H/D exchange) for measuring the stability of proteins in a rapid, high-throughput fashion. The method uses hydrogen exchange to estimate the stability of microgram quantities of unpurified protein extracts by using matrix-assisted laser desorption/ionization MS. The stabilities of maltose binding protein and monomeric lambda repressor variants determined by SUPREX agree well with stability data obtained from conventional CD denaturation of purified protein. The method also can detect the change in stability caused by the binding of maltose to maltose binding protein. The results demonstrate the precision of the method over a wide range of stabilities.
Duke Scholars
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Related Subject Headings
- Viral Regulatory and Accessory Proteins
- Viral Proteins
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
- Repressor Proteins
- Recombinant Fusion Proteins
- Maltose-Binding Proteins
- Maltose
- DNA-Binding Proteins
- Carrier Proteins
- Bacteriophage lambda
Citation
Published In
DOI
ISSN
Publication Date
Volume
Issue
Start / End Page
Location
Related Subject Headings
- Viral Regulatory and Accessory Proteins
- Viral Proteins
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
- Repressor Proteins
- Recombinant Fusion Proteins
- Maltose-Binding Proteins
- Maltose
- DNA-Binding Proteins
- Carrier Proteins
- Bacteriophage lambda