5UYO

Solution NMR structure of the de novo mini protein HEEH_rd4_0097

Summary for 5UYO

• Entry DOI: 10.2210/pdb5uyo/pdb
• NMR Information: BMRB: 30249
• Descriptor: HEEH_rd4_0097 (1 entity in total)
• Functional Keywords: de novo design, helix-strand-strand-helix, mini protein, de novo protein
• Biological source: Escherichia coli
• Total number of polymer chains: 1
• Total formula weight: 7469.42

Authors

Lemak, A.,Rocklin, G.J.,Houliston, S.,Carter, L.,Chidyausiku, T.M.,Baker, D.,Arrowsmith, C.H. (deposition date: 2017-02-24, release date: 2017-07-26, Last modification date: 2024-05-15)

Primary citation

Rocklin, G.J.,Chidyausiku, T.M.,Goreshnik, I.,Ford, A.,Houliston, S.,Lemak, A.,Carter, L.,Ravichandran, R.,Mulligan, V.K.,Chevalier, A.,Arrowsmith, C.H.,Baker, D.
Global analysis of protein folding using massively parallel design, synthesis, and testing.
Science, 357:168-175, 2017

PubMed Abstract: Proteins fold into unique native structures stabilized by thousands of weak interactions that collectively overcome the entropic cost of folding. Although these forces are "encoded" in the thousands of known protein structures, "decoding" them is challenging because of the complexity of natural proteins that have evolved for function, not stability. We combined computational protein design, next-generation gene synthesis, and a high-throughput protease susceptibility assay to measure folding and stability for more than 15,000 de novo designed miniproteins, 1000 natural proteins, 10,000 point mutants, and 30,000 negative control sequences. This analysis identified more than 2500 stable designed proteins in four basic folds-a number sufficient to enable us to systematically examine how sequence determines folding and stability in uncharted protein space. Iteration between design and experiment increased the design success rate from 6% to 47%, produced stable proteins unlike those found in nature for topologies where design was initially unsuccessful, and revealed subtle contributions to stability as designs became increasingly optimized. Our approach achieves the long-standing goal of a tight feedback cycle between computation and experiment and has the potential to transform computational protein design into a data-driven science.

PubMed: 28706065
DOI: 10.1126/science.aan0693

Experimental method

SOLUTION NMR