Probing the structure and stability of a hybrid protein: the human-E. coli thioredoxin chimera.


The structure and stability of a hybrid protein composed of N-terminal human and C-terminal E. coli thioredoxin domains were investigated by NMR, fluorescence, and circular dichroism spectroscopy. We demonstrate that the chimeric protein is correctly folded and exhibits the common thioredoxin architecture. However, the stability of the hybrid protein toward thermal and chemical denaturation is clearly reduced when compared with both parent proteins. Altogether, our data indicate that the interface between the two folding units of thioredoxin is tolerant toward changes in exact interdigitation of side chains, allowing for the formation of the unique overall thioredoxin fold. Further, the gene encoding the human-E. coli chimera was tested in vivo whether it supports the assembly of filamentous phages. No complementation of a thioredoxin-deficient E. coli mutant for the replication of the phages M13 or fd was observed, suggesting that parts of the overall protein structure in the N-terminal domain are crucial for this activity. Study holds ProTherm entries: 11712, 11713 Extra Details: chimeric protein; hybrid; thioredoxin fold; activity

Submission Details

ID: jwvKeGNf

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:42 p.m.

Version: 1

Publication Details
Louis JM;Georgescu RE;Tasayco ML;Tcherkasskaya O;Gronenborn AM,Biochemistry (2001) Probing the structure and stability of a hybrid protein: the human-E. coli thioredoxin chimera. PMID:11551217
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
100.0 Thioredoxin 1 P0AA27 THIO_ECO57
100.0 Thioredoxin 1 P0AA26 THIO_ECOL6
100.0 Thioredoxin 1 P0AA25 THIO_ECOLI
100.0 Thioredoxin 1 P0AA29 THIO_SALTI
100.0 Thioredoxin 1 P0AA28 THIO_SALTY
100.0 Thioredoxin 1 P0AA30 THIO_SHIFL
100.0 Thioredoxin P10599 THIO_HUMAN
97.1 Thioredoxin P29451 THIO_MACMU
99.1 Thioredoxin Q5R9M3 THIO_PONAB
96.2 Thioredoxin Q9BDJ3 THIO_CALJA
94.3 Thioredoxin P82460 THIO_PIG
93.3 Thioredoxin O97680 THIO_BOVIN
92.4 Thioredoxin P50413 THIO_SHEEP
90.5 Thioredoxin O97508 THIO_HORSE
90.3 Thioredoxin P11232 THIO_RAT