AI Article Synopsis
- A new combinatorial selective labeling (CSL) method has been developed for assigning backbone amide NMR resonances, useful for identifying protein-ligand interactions.
- The CSL technique utilizes a variety of samples with different labeled amino acid patterns, allowing for simultaneous assignment of multiple amino acid combinations through peak intensity analysis in HSQC and 2D HNCO spectra.
- Demonstrated on the 27 kDa protein GFP, this method is quick, cost-effective, and can be easily automated for streamlined backbone assignment.
Article Abstract
A combinatorial selective labeling (CSL) method is presented for the assignment of backbone amide NMR resonances, which has a particular application in the identification of protein-ligand interaction sites. The method builds on the dual amino acid selective labeling technique. In the CSL method a number of different samples are produced, each with a different pattern of labeled amino acids. By analyzing peak intensities in HSQC and 2D HNCO spectra of these samples, a large number of combinations of amino acid pairs can be simultaneously assigned. We demonstrate the method on the 27 kDa protein GFP. The samples can be produced rapidly and cost-effectively in a commercially available in vitro translation system. The method greatly simplifies the process of backbone assignment and would be very straightforward to automate.
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|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1800590 | PMC |
| http://dx.doi.org/10.1021/ja039601r | DOI Listing |
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