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The Escherichia coli-derived thymosin ?4 concatemer promotes cell proliferation and healing wound in mice.

ABSTRACT: Thymosin ?4 (T?4) is one of the most promising thymosins for future clinical applications, and it is anticipated that commercial demand for T?4 will increase. In order to develop a new approach to produce recombinant T?4, a 168 bp DNA (termed T?4) was designed based on the T?4 protein sequence and used to express a 4 × T? 4 concatemer (four tandem copies of T?4, termed 4 × T?4) together with a histidine tag (6 × His) in E. coli (strain BL21). SDS-PAGE and western blot analysis were used to confirm that a recombinant 4 × T?4 protein of the expected size (30.87 kDa) was produced following the induction of the bacterial cultures with isopropyl ?-D-thiogalactoside (IPTG). The E. coli-derived 4 × T?4 was purified by Ni-NTA resin, and its activities were examined with regard to both stimulating proliferation of the mice spleen cells in vitro and in vivo wound healing. The results demonstrate that these activities of the E. coli-derived recombinant 4 × T?4 were similar or even better than existing commercially obtained T?4. This production strategy therefore represents a potentially valuable approach for future commercial production of recombinant T?4.

SUBMITTER: Wang X

PROVIDER: S-EPMC3671520 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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The Escherichia coli-derived thymosin β4 concatemer promotes cell proliferation and healing wound in mice.

Wang Xiaolei X Yang Guihua G Li Shanshuang S Gao Meifeng M Zhao Pangfeng P Zhao Lingxia L

BioMed research international 20130519

Thymosin β4 (Tβ4) is one of the most promising thymosins for future clinical applications, and it is anticipated that commercial demand for Tβ4 will increase. In order to develop a new approach to produce recombinant Tβ4, a 168 bp DNA (termed Tβ4) was designed based on the Tβ4 protein sequence and used to express a 4 × Tβ 4 concatemer (four tandem copies of Tβ4, termed 4 × Tβ4) together with a histidine tag (6 × His) in E. coli (strain BL21). SDS-PAGE and western blot analysis were used to confi ...[more]

PMID: 23762829

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Project description:This study investigates the effects of environmentally-relevant concentrations of fluoxetine (FLX, commercial name: Prozac) on wound healing. Pollution of water systems with pharmaceutical and care products, including antidepressants such as FLX and other selective serotonin reuptake inhibitors, is a growing environmental concern. Environmentally-relevant FLX concentrations are known to impact physiological functions and behaviour of aquatic animals, however, the effects of exposure on humans are currently unknown. Using a combination of human skin biopsies and keratinocyte cell lines, we show that exposure to environmental FLX enhances wound closure. We show dose-dependent increases in wound closure with FLX concentrations from 125 ng/l. We demonstrate that the mechanisms underlying enhanced wound closure are increased cell proliferation and serotonin signalling. Transcriptomic analysis revealed 350 differentially expressed genes after exposure. Downregulated genes were enriched in pathways related to mitochondrial function and metabolism, while upregulated genes were associated with cell proliferation and tissue morphogenesis. Kinase profiling showed altered phosphorylation of kinases related to the MAPK pathway. Consistent with this, phosphoproteomics analyses identified 235 differentially phosphorylated proteins after exposure, with enriched GO terms related to cell cycle, division, and protein biosynthesis. These findings collectively show that exposure to environmental FLX promotes wound healing through modulation of serotonin signalling, gene expression and protein phosphorylation, leading to enhanced cell proliferation. Our results open the door for further research to investigate the effects of environmental FLX on cell metabolism and proliferation, and clinical applications of FLX in the setting of wound healing. We argue that there is an urgent need to develop models that can inform on the risks, and potential benefits, of environmental pollution with selective serotonin reuptake inhibitors.

Dataset Information

The Escherichia coli-derived thymosin ?4 concatemer promotes cell proliferation and healing wound in mice.

Publications

The Escherichia coli-derived thymosin β4 concatemer promotes cell proliferation and healing wound in mice.

Similar Datasets

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