We have previously reported a phage display method for the identification of protein domains on a genome-wide scale (shotgun proteolysis). Here we present the solution structure of a fragment of the Escherichia coli membrane protein yrfF, as identified by shotgun proteolysis, and determined by NMR spectroscopy. Despite the absence of computational predictions, the fragment formed a well-defined beta-barrel structure, distantly falling within the OB-fold classification. Our results highlight the potential of high-throughput experimental approaches for the identification of protein domains for structural studies.
Allen, M. D., Christie, M., Jones, P., Porebski, B. T., Roome, B., Freund, S. M. V., … Christ, D. (2015). Solution structure of a soluble fragment derived from a membrane protein by shotgun proteolysis. Protein Engineering, Design and Selection, 28(10), 445–450. https://doi.org/10.1093/protein/gzv021