Improvement of the recombinant human coagulation factor IX expression by co-expression of a novel transcript of Drosophila γ carboxylase in a human cell line.
Biotechnol Lett
; 42(11): 2147-2156, 2020 Nov.
Article
de En
| MEDLINE
| ID: mdl-32514789
ABSTRACT
OBJECTIVE:
Mammalian cells as the main host for production of human proteins are incapable of complete γ-carboxylation of over-expressed Vitamin K Dependent (VKD) proteins. The Drosophila γ-glutamyl carboxylase (DγC) has been shown to be more efficient than its human counterpart in γ-carboxylation of human substrates, in vitro. Considering the Drosophila γ-carboxylase (DγC) efficiency, in comparison with its human counterpart, for recognition and γ-carboxylation of a human substrate in vitro, we were determined to study the effect of the DγC on the hFIX expression in a mammalian cell line. With this aim, we examined co-expression of the DγC with the hFIX, in a human cell line.RESULTS:
While the co-expression of a complete DγC cDNA reduced the hFIX expression, a truncated form of DγC could improve both the expression level (up to 1211 ng/106 cells/ml on the 4th day of post-transfection) and carboxylation of the expressed hFIX, significantly (p < 0.009).CONCLUSIONS:
Our findings provided evidences for potential of a partial fragment of the DγC for improvement of the γ-carboxylation of a human substrate in a mammalian cell. Our experimental data, in accordance with in silico analysis suggested that the DγC C-terminal fragment, with the advantage of a Kozak-like element has the potential of being expressed as a separate internal translation unit, to generate a peptide with appropriate γ-carboxylase activity.Mots clés
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Facteur IX
/
Carbon-carbon ligases
/
Drosophila
/
Mutation
Limites:
Animals
/
Humans
Langue:
En
Journal:
Biotechnol Lett
Année:
2020
Type de document:
Article
Pays d'affiliation:
Iran