Improvement of thermostability of recombinant collagen-like protein by incorporating a foldon sequence.
Collagen is a popular biomaterial in many specific biological interactions as well as a structural element. In this work, the recombinant collagen-like proteins were synthesized using Escherichia coli expression system. A foldon sequence, GYIPEAPRDGQAYVRKDG EWVLLSTFL, derived from the native T4 phage fibritin was incorporated at the C-terminal of collagen-like protein molecules to stabilize the triple helix formed in the proteins. The differential scanning calorimetry and thermogravimetric analysis measurements showed that the thermostability of the recombinant collagen-like proteins was significantly improved when compared with those without the foldon sequence at the C-terminal. Fourier transform infrared and scanning electron microscopy observations indicated that the collagen-like proteins forms the triple helix structure and prefer to aggregate as fibrils, same as the native collagen. Moreover, the mice fibroblasts L929 cells could attach and grew very well on the recombinant collage-like proteins. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that the cell biocompatibility of collagen-like proteins produced in this work was even better than that of native collagen, suggesting that the collagen-like proteins may be a satisfactory candidate for the future applications as a biomaterial.
Du
Chunling
C
Key Laboratory of Advanced Textile Materials and Manufacturing Technology of Ministry of Education, College of Materials and Textile, Zhejiang Sci-Tech University, Hangzhou 310018, People's Republic of China.
Wang
Mingqi
M
Liu
Jinying
J
Pan
Mingli
M
Cai
Yurong
Y
Yao
Juming
J
D000818:Animals; D017122:Bacteriophage T4 / Q000737:chemistry* / Q000235:genetics; D001672:Biocompatible Materials; D003094:Collagen / Q000737:chemistry* / Q000235:genetics; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D005347:Fibroblasts; D051379:Mice; D008855:Microscopy, Electron, Scanning; D015202:Protein Engineering; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012995:Solubility; D017550:Spectroscopy, Fourier Transform Infrared; D014764:Viral Proteins / Q000378:metabolism
D001672:Biocompatible Materials; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; C057839:fibritin protein, Enterobacteria phage T4; D003094:Collagen