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Comparison of the N-linked glycosylation of human β1,3-N-acetylglucosaminyltransferase 2 expressed in insect cells and silkworm larvae

Paper ID Volume ID Publish Year Pages File Format Full-Text
24723 43532 2009 7 PDF Available
Title
Comparison of the N-linked glycosylation of human β1,3-N-acetylglucosaminyltransferase 2 expressed in insect cells and silkworm larvae
Abstract

N-Glycosylation of human β1,3N-acetylglucosaminyltransferase 2 (β3GnT2) is essential for its biological function. β3GnT2 fused to GFPuv (GFPuv–β3GnT2) was produced by non-virus expression systems in stably transformed insect cells and silkworm larvae using a recombinant BmNPV bacmid, and purified for analysis of N-glycosylation. The N-glycan structure of β3GnT2 was identified by glycoamidase A digestion, labeling with 2-aminopyridine (PA), and HPLC mapping. The paucimannosidic N-glycan structure (73.2%) was predominant in stably transformed Trichoplusia ni cells. In contrast, N-glycan with Gal (21.3%) and GlcNAc (16.2%) terminal residues linked to Manα(1,3) branch were detected on β3GnT2 expressed in silkworm larvae. The presence of terminal Gal and bisecting GlcNAc residues such as Galβ1, 4GlcNAcβ1, 2Manα1,3(GlcNAcβ1,4)(Manα1,6)Manβ1, 4GlcNAc is not typical structure for lepidopteran insect N-glycosylation. Although allergenic α1,3-fucose residues have been found in T. ni cells, only α1,6-fucose residues were attached to the β3GnT2 glycan in silkworm larvae. Therefore, silkworm larvae might be a useful host for producing human glycoproteins.

Keywords
Silkworm larvae; BmNPV bacmid; Insect cell; β1,3N-Acetylglucosaminyltransferase 2; N-Glycosylation
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Comparison of the N-linked glycosylation of human β1,3-N-acetylglucosaminyltransferase 2 expressed in insect cells and silkworm larvae
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 143, Issue 1, 10 August 2009, Pages 27–33
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us