| RRC ID |
35610
|
| 著者 |
Singh P, Sharma L, Kulothungan SR, Adkar BV, Prajapati RS, Ali PS, Krishnan B, Varadarajan R.
|
| タイトル |
Effect of signal peptide on stability and folding of Escherichia coli thioredoxin.
|
| ジャーナル |
PLoS One
|
| Abstract |
The signal peptide plays a key role in targeting and membrane insertion of secretory and membrane proteins in both prokaryotes and eukaryotes. In E. coli, recombinant proteins can be targeted to the periplasmic space by fusing naturally occurring signal sequences to their N-terminus. The model protein thioredoxin was fused at its N-terminus with malE and pelB signal sequences. While WT and the pelB fusion are soluble when expressed, the malE fusion was targeted to inclusion bodies and was refolded in vitro to yield a monomeric product with identical secondary structure to WT thioredoxin. The purified recombinant proteins were studied with respect to their thermodynamic stability, aggregation propensity and activity, and compared with wild type thioredoxin, without a signal sequence. The presence of signal sequences leads to thermodynamic destabilization, reduces the activity and increases the aggregation propensity, with malE having much larger effects than pelB. These studies show that besides acting as address labels, signal sequences can modulate protein stability and aggregation in a sequence dependent manner.
|
| 巻・号 |
8(5)
|
| ページ |
e63442
|
| 公開日 |
2013-1-1
|
| DOI |
10.1371/journal.pone.0063442
|
| PII |
PONE-D-12-30695
|
| PMID |
23667620
|
| PMC |
PMC3646739
|
| MeSH |
Amino Acid Sequence
Anilino Naphthalenesulfonates / metabolism
Buffers
Calorimetry, Differential Scanning
Chromatography, Gel
Circular Dichroism
Electrophoresis, Polyacrylamide Gel
Escherichia coli / metabolism*
Guanidine / pharmacology
Insulin / metabolism
Molecular Sequence Data
Protein Denaturation / drug effects
Protein Folding* / drug effects
Protein Refolding / drug effects
Protein Sorting Signals*
Protein Stability / drug effects
Protein Structure, Quaternary
Proteolysis / drug effects
Recombinant Fusion Proteins / metabolism
Spectrometry, Fluorescence
Temperature
Thioredoxins / chemistry*
Thioredoxins / metabolism*
|
| IF |
2.74
|
| 引用数 |
26
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| 原核生物(大腸菌) |
|
