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Molecular assembly of recombinant chicken type II collagen in the yeast Pichia pastoris

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Abstract

Effective treatment of rheumatoid arthritis can be mediated by native chicken type II collagen (nCCII), recombinant peptide containing nCCII tolerogenic epitopes (CTEs), or a therapeutic DNA vaccine encoding the full-length CCOL2A1 cDNA. As recombinant CCII (rCCII) might avoid potential pathogenic virus contamination during nCCII preparation or chromosomal integration and oncogene activation associated with DNA vaccines, here we evaluated the importance of propeptide and telopeptide domains on rCCII triple helix molecular assembly. We constructed pC- and pN-procollagen (without N- or C-propeptides, respectively) as well as CTEs located in the triple helical domain lacking both propeptides and telopeptides, and expressed these in yeast Pichia pastoris host strain GS115 (his4, Mut+) simultaneously with recombinant chicken prolyl-4-hydroxylase α and β subunits. Both pC- and pN-procollagen monomers accumulated inside P. pastoris cells, whereas CTE was assembled into homotrimers with stable conformation and secreted into the supernatants, suggesting that the large molecular weight pC-or pN-procollagens were retained within the endoplasmic reticulum whereas the smaller CTEs proceeded through the secretory pathway. Furthermore, resulting recombinant chicken type II collagen pCα1(II) can induced collagen-induced arthritis (CIA) rat model, which seems to be as effective as the current standard nCCII. Notably, protease digestion assays showed that rCCII could assemble in the absence of C- and N-propeptides or telopeptides. These findings provide new insights into the minimal structural requirements for rCCII expression and folding.

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Authors and Affiliations

  1. Department of Immunology and National Center for Biomedicine Analysis, Beijing 307 Hospital Affiliated to Academy of Military Medical Sciences, Beijing, 100071, China

    Caixia Xi, Nan Liu, Fei Liang, Xiao Zhao, Juan Long, Fang Yuan, Song Yun, Yuying Sun & Yongzhi Xi

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  1. Caixia Xi
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  2. Nan Liu
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  3. Fei Liang
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  4. Xiao Zhao
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  5. Juan Long
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  6. Fang Yuan
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  7. Song Yun
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  8. Yuying Sun
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  9. Yongzhi Xi
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Correspondence to Yongzhi Xi.

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Xi, C., Liu, N., Liang, F. et al. Molecular assembly of recombinant chicken type II collagen in the yeast Pichia pastoris. Sci. China Life Sci. 61, 815–825 (2018). https://doi.org/10.1007/s11427-017-9219-4

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  • DOI: https://doi.org/10.1007/s11427-017-9219-4

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