Dataset Information

Construction and Activity Analyses of Single Functional Mouse Peroxiredoxin 6 (Prdx6).

ABSTRACT:

Introduction

Peroxiredoxin 6 (Prdx6) is a bifunctional protein with glutathione peroxidase activity and phospholipase A2 activity. Previous studies have shown a significant positive correlation between the intracellular survival ability of Brucella and Prdx6. Here, the Prdx6 enzyme with a single activity was constructed to facilitate study of the relationship between the single function of Prdx6 and Brucella infection.

Material and methods

The target open reading frame (ORF) DNAs of Prdx6 with a single active centre were prepared using gene splicing by overlap extension PCR (SOE-PCR), and the recombinant eukaryotic expression plasmids inserted by Prdx6 with the single activity centre were constructed and transfected into murine Raw264.7 macrophages. The glutathione peroxidase activity and phospholipase A2 activity of the constructed Prdx6 were examined.

Results

The core centres (Ser³² and Cys⁴⁷) of Prdx6 were successfully mutated by changing the 94^th nucleotide from T to G and the 140^th nucleotide from G to C in the two enzyme activity cores, respectively. The constructed recombinant plasmids of Prdx6 with the single active centre were transfected into murine macrophages showing the expected single functional enzyme activity, which MJ33 or mercaptosuccinate inhibitors were able to inhibit.

Conclusion

The constructed mutants of Prdx6 with the single activity cores will be a benefit to further study of the biological function of Prdx6 with different enzyme activity.

SUBMITTER: Wang LL

PROVIDER: S-EPMC6458554 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

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Publications

Construction and Activity Analyses of Single Functional Mouse Peroxiredoxin 6 (Prdx6).

Wang Lu-Lu LL Lu Shi-Ying SY Hu Pan P Fu Bao-Quan BQ Li Yan-Song YS Zhai Fei-Fei FF Ju Dan-Di DD Zhang Shi-Jun SJ Su Bing B Zhou Yu Y Liu Zeng-Shan ZS Ren Hong-Lin HL

Journal of veterinary research 20190322 1

<h4>Introduction</h4>Peroxiredoxin 6 (Prdx6) is a bifunctional protein with glutathione peroxidase activity and phospholipase A2 activity. Previous studies have shown a significant positive correlation between the intracellular survival ability of <i>Brucella</i> and Prdx6. Here, the Prdx6 enzyme with a single activity was constructed to facilitate study of the relationship between the single function of Prdx6 and <i>Brucella</i> infection.<h4>Material and methods</h4>The target open reading fra ...[more]

PMID: 30989141

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Dataset Information

Construction and Activity Analyses of Single Functional Mouse Peroxiredoxin 6 (Prdx6).

Introduction

Material and methods

Results

Conclusion

Publications

Construction and Activity Analyses of Single Functional Mouse Peroxiredoxin 6 (Prdx6).

Similar Datasets

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