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Guanidinium derivatives bind preferentially and trigger long-distance conformational changes in an engineered T4 lysozyme.

Yousef MS, Bischoff N, Dyer CM, Baase WA, Matthews BW
Protein Sci 15 853-61 (2006)
Related entries: 1oyu, 1t8a, 1t97, 261l, 262l, 2f32, 2f47

Cited: 2 times
EuropePMC logo PMID: 16600969

Abstract

The binding of guanidinium ion has been shown to promote a large-scale translation of a tandemly duplicated helix in an engineered mutant of T4 lysozyme. The guanidinium ion acts as a surrogate for the guanidino group of an arginine side chain. Here we determine whether methyl- and ethylguanidinium provide better mimics. The results show that addition of the hydrophobic moieties to the ligand enhances the binding affinity concomitant with reduction in ligand solubility. Crystallographic analysis confirms that binding of the alternative ligands to the engineered site still drives the large-scale conformational change. Thermal analysis and NMR data show, in comparison to guanidinium, an increase in protein stability and in ligand affinity. This is presumably due to the successive increase in hydrophobicity in going from guanidinium to ethylguanidinium. A fluorescence-based optical method was developed to sense the ligand-triggered helix translation in solution. The results are a first step in the de novo design of a molecular switch that is not related to the normal function of the protein.

Articles citing this publication (2)

  1. Studies on the functional significance of a C-terminal S-shaped motif in human arginase type I: essentiality for cooperative effects. García D, Uribe E, Lobos M, Orellana MS, Carvajal N. Arch Biochem Biophys 481 16-20 (2009)
  2. Molecular dynamics simulations of an engineered T4 lysozyme exclude helix to sheet transition, and provide insights into long distance, intra-protein switchable motion. Biggers L, Elhabashy H, Ackad E, Yousef MS. Protein Sci 29 542-554 (2020)


Related citations provided by authors (3)

  1. Use of sequence duplication to engineer a ligand-triggered, long-distance molecular switch in T4 lysozyme.. Yousef MS, Baase WA, Matthews BW Proc Natl Acad Sci U S A 101 11583-6 (2004)
  2. Structural characterization of an engineered tandem repeat contrasts the importance of context and sequence in protein folding.. Sagermann M, Baase WA, Matthews BW Proc Natl Acad Sci U S A 96 6078-83 (1999)
  3. Long-distance conformational changes in a protein engineered by modulated sequence duplication.. Sagermann M, Gay L, Matthews BW Proc Natl Acad Sci U S A 100 9191-5 (2003)

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