The alpha subunit of tryptophan synthase from Escherichia coli has been previously shown to contain residual structure at 5 M urea, conditions where the secondary structure is entirely disrupted and the tyrosine residues are exposed to solvent [Saab-Rincón, G., Froebe, C. L., & Matthews, C. R. (1993) Biochemistry 32, 13981-13990]. The residual structure can be monitored by one-dimensional NMR spectroscopy studies of histidine 92 whose C epsilon proton is sensitive to the slow exchange between this form and the unfolded protein. The temperature dependence of the cooperative urea-induced unfolding transition between intermediate and unfolded forms demonstrates that this process involves negative values for both the enthalpy and entropy changes at 25 degrees C. The effects of replacements of several nonpolar side chains adjacent to histidine 92 on the slopes and midpoints of the unfolding transition curve show that these side chains participate in the residual structure. A 15-residue peptide spanning histidine 92 and the mutated residues, however, is not sufficient to define this structure. These results demonstrate that the residual structure in the alpha subunit is stabilized by the hydrophobic effect and may involve side chains which are distant in sequence to histidine 92. Study holds ProTherm entries: 4910, 4911, 4912, 4913, 4914, 4915, 4916, 4917, 4918, 4919, 4920, 4921, 4922 Extra Details: additive : EDTA(0.2 mM), hydrophobic effect
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:27 p.m.
|Number of data points||25|
|Proteins||Tryptophan synthase alpha chain ; Tryptophan synthase alpha chain|
|Assays/Quantities/Protocols||Experimental Assay: Cm temp:5.0 C ; Experimental Assay: dG_H2O temp:5.0 C ; Experimental Assay: Cm temp:10.0 C ; Experimental Assay: dG_H2O temp:10.0 C ; Experimental Assay: Cm temp:15.0 C ; Experimental Assay: dG_H2O temp:15.0 C ; Experimental Assay: Cm temp:20.0 C ; Experimental Assay: dG_H2O temp:20.0 C ; Experimental Assay: Cm temp:25.0 C ; Experimental Assay: dG_H2O temp:25.0 C|
|Libraries||Mutations for sequence MERYESLFAQLKERKEGAFVPFVTLGDPGIEQSLKIIDTLIEAGADALELGIPFSDPLADGPTIQNATLRAFAAGVTPAQCFEMLALIRQKHPTIPIGLLMYANLVFNKGIDEFYAQCEKVGVDSVLVADVPVEESAPFRQAALRHNVAPIFICPPNADDDLLRQIASYGRGYTYLLSRAGVTGAENRAALPLNHLVAKLKEYNAAPPLQGFGISAPDQVKAAIDAGAAGAISGSAIVKIIEQHINEPEKMLAALKVFVQPMKAATRS|