|
KMID : 0613820220320020135
|
|
Journal of Life Science
2022 Volume.32 No. 2 p.135 ~ p.141
|
|
|
Cell Migration and Wound Healing Activities of Recombinant Thymosin β-4 Expressed in Escherichia coli
|
|
Hong Kyo-Chang
Choi Yung-Hyun
Kim Gun-Do
Cha Hee-Jae
Jeon Sung-Jong
Nam Soo-Wan
|
|
|
Abstract
|
|
|
|
Thymosin β-4 (TB4) is a small peptide composed of 43 amino acids. To obtain sufficient biologically active mouse TB4 economically, we cloned and overexpressed this gene in an Escherichia coli system. With the isopropyl β-D-1-thiogalactopyranoside induction of the E. coli transformant, TB4 fusion protein with intein- and chitin-binding domain was successfully expressed in the soluble fraction within the E. coli cell. The TB4-intein?chitin-binding domain fusion protein was purified from the soluble fraction of E. coli cell lysate. The affinity chromatography with chitin beads and dithiothreitol-mediated intein self-cleavage reaction releases the TB4 peptide into the stripping solution. Sodium dodecyl sulphate?polyacrylamide gel electrophoresis and Western blot analyses were used to confirm that the recombinant TB4 peptide was produced with the expected size of 5 kDa. We found that the recombinant TB4 stimulated cell migration in the transwell plate chamber assay. After 18 hr of the treatment of the recombinant TB4 with 1 ng/ml concentration, the migration of the HT1080 cell was increased by 20% compared with that of the chemically synthesized TB4. The recombinant TB4 was also observed to promote the healing of a wound area in C57BL/6 mice by as high as 35% compared with that of the chemically synthesized TB4. These results suggest that the recombinant TB4 has better biological activity for cell migration and wound healing than that of the chemically synthesized TB4 peptide.
|
|
|
KEYWORD
|
|
|
Cell migration, E. coli, Thymosin beta-4, wound healing
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|