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Title 

High-level expression of human glycosyltransferases in insect cells as biochemically active form

Authors 

H G KimS M YangY C LeeSu Il DoI S ChungJ M Yang

Publisher 

Elsevier

Issue Date 

2003

Citation 

Biochemical and Biophysical Research Communications, vol. 305, no. 3, pp. 488-493

Keywords 

galactosyltransferaseheterologous expressionIgM signal peptideinsect cellprotein Asialyltransferaseanimal cellcell culturecell lineexpression vector

Abstract 

cDNAs, encoding human β1,4-galactosyltransferase (hGalT I, EC 2.4.1.22), human Galβ1,3(4)-GlcNAc α2,3-sialyltransferase (hST3GalIII, EC 2.4.99), and human Galβ1,4-GlcNAc α2,6-sialyltransferase (hST6Gal I, EC 2.4.99.1), were cloned from human cell lines. In order to express these glycosyltransferases as secreted form in insect cells, cDNAs were inserted into a novel baculovirus transfer vector equipped with the mouse IgM signal peptide and IgG binding domain of the Staphylococcus aureus protein A as an N-terminal fusion partner. About 14mg hGalT I, 8mg hST3GalIII, and 6.4mg hST6Gal I were purified from 1liter of recombinant baculovirus infected insect cell culture media. The specific activities of recombinant hGalT I and hST6Gal I were determined as 0.65 and 1.6U/mg protein, respectively. These results indicated that the recombinant hGalT I and hST6Gal I retained enzyme activities at similar level to those of the authentic one although they were fused with the IgG binding domain at the N-terminus. Taken together, the mouse IgM signal peptide and IgG binding domain of the protein A could be efficiently used as an N-terminus fusion partner for the over-expression of heterologous proteins in insect cells.

ISSN 

0006-291X

Link 

http://dx.doi.org/10.1016/S0006-291X(03)00795-2

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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