A single point mutation (Glu85Arg) increases the stability of the thioredoxin from Escherichia coli.
Abstract
Glu85 in the Escherichia coli thioredoxin, which is localized in the loop between beta4 and beta5, was substituted with the Arg present in the corresponding position in Bacillus acidocaldarius thioredoxin. This suggested that it could play an important role in the structure and thermostability of this protein owing to its involvement in numerous interactions. The effects of the mutation on the biophysical properties were analysed by circular dichroism, spectrofluorimetry and limited proteolysis, supported by molecular dynamics data. As modelling predicted, an increase in stability for E85R due to additional H-bonds between the beta5 and alpha4 regions was observed. Study holds ProTherm entries: 10820 Extra Details: molecular dynamic simulations; protein engineering;,site-directed mutagenesis; thermostability; thioredoxin
Submission Details
ID: XUqK7HdR3
Submitter: Connie Wang
Submission Date: April 24, 2018, 8:41 p.m.
Version: 1
Publication Details
Pedone E;Saviano M;Rossi M;Bartolucci S,Protein Eng. (2001) A single point mutation (Glu85Arg) increases the stability of the thioredoxin from Escherichia coli. PMID:11391017Additional Information
Study Summary
| Number of data points | 1 |
| Proteins | Thioredoxin 1 ; Thioredoxin 1 |
| Unique complexes | 1 |
| Assays/Quantities/Protocols | Experimental Assay: Tm |
| Libraries | Mutations for sequence SDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDEIADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFLDANLA |
Structure view and single mutant data analysis
Study data
| Colors: | D | E | R | H | K | S | T | N | Q | A | V | I | L | M | F | Y | W | C | G | P |
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Data Distribution
Studies with similar sequences (approximate matches)
Correlation with other assays (exact sequence matches)
Relevant PDB Entries
| Structure ID | Release Date | Resolution | Structure Title |
|---|---|---|---|
| 5IKN | 2016-03-03T00:00:00+0000 | 4.8 | Crystal Structure of the T7 Replisome in the Absence of DNA |
| 5E4W | 2015-10-07T00:00:00+0000 | 2.8 | Crystal structure of cpSRP43 chromodomains 2 and 3 in complex with the Alb3 tail |
| 3DXB | 2008-07-24T00:00:00+0000 | 2.2 | Structure of the UHM domain of Puf60 fused to thioredoxin |
| 1T8E | 2004-05-12T00:00:00+0000 | 2.54 | T7 DNA Polymerase Ternary Complex with dCTP at the Insertion Site. |
| 6P7E | 2019-06-05T00:00:00+0000 | 3.0 | Structure of T7 DNA Polymerase Bound to a Primer/Template DNA and a Peptide that Mimics the C-terminal Tail of the Primase-Helicase |
| 2EIR | 2007-03-13T00:00:00+0000 | 2.5 | Design of Disulfide-linked Thioredoxin Dimers and Multimers Through Analysis of Crystal Contacts |
| 2EIO | 2007-03-13T00:00:00+0000 | 2.6 | Design of Disulfide-linked Thioredoxin Dimers and Multimers Through Analysis of Crystal Contacts |
| 1ZCP | 2005-04-12T00:00:00+0000 | 2.3 | Crystal Structure of a catalytic site mutant E. coli TrxA (CACA) |
| 1X9S | 2004-08-24T00:00:00+0000 | 2.7 | T7 DNA polymerase in complex with a primer/template DNA containing a disordered N-2 aminofluorene on the template, crystallized with dideoxy-CTP as the incoming nucleotide. |
| 2EIQ | 2007-03-13T00:00:00+0000 | 1.9 | Design of Disulfide-linked Thioredoxin Dimers and Multimers Through Analysis of Crystal Contacts |