scholarly journals Glycosylation in Lepidopteran insect cells: identification of a β1→4-N-acetylgalactosaminyltransferase involved in the synthesis of complex-type oligosaccharide chains

Glycobiology ◽  
1996 ◽  
Vol 6 (2) ◽  
pp. 157-164 ◽  
Author(s):  
Irma van Die ◽  
Angelique van Tetering ◽  
Hans Bakker ◽  
Dirk H. van den Eijnden ◽  
David H. Joziasse
Keyword(s):  
Insect Cells ◽  
Complex Type ◽  
Lepidopteran Insect

scholarly journals Site-specific N-glycosylation Characterization of Recombinant SARS-CoV-2 Spike Proteins

2020 ◽  
pp. mcp.RA120.002295 ◽  
Author(s):  
Yong Zhang ◽  
Wanjun Zhao ◽  
Yonghong Mao ◽  
Yaohui Chen ◽  
Shisheng Wang ◽  
...  
Keyword(s):  
Insect Cells ◽  
Host Cells ◽  
Protein Subunit ◽  
Genome Sequences ◽  
Complex Type ◽  
Site Specific ◽  
Differential Processing ◽  
S Protein ◽  
Shed Light

The glycoprotein spike (S) on the surface of SARS-CoV-2 is a determinant for viral invasion and host immune response. Herein, we characterized the site-specific N-glycosylation of S protein at the level of intact glycopeptides. All 22 potential N-glycosites were identified in the S-protein protomer and were found to be preserved among the 753 SARS-CoV-2 genome sequences. The glycosites exhibited glycoform heterogeneity as expected for a human cell-expressed protein subunit. We identified masses that correspond to 157 N-glycans, primarily of the complex type. In contrast, the insect cell-expressed S protein contained 38 N-glycans, completely of the high-mannose type. Our results revealed that the glycan types were highly determined by the differential processing of N-glycans among human and insect cells, regardless of the glycosites’ location. Moreover, the N-glycan compositions were conserved among different sizes of subunits. Our study indicate that the S protein N-glycosylation occurs regularly at each site, albeit the occupied N-glycans were diverse and heterogenous. This N-glycosylation landscape and the differential N-glycan patterns among distinct host cells are expected to shed light on the infection mechanism and present a positive view for the development of vaccines and targeted drugs.

ECDYSONE-INDUCIBLE FOREIGN GENE EXPRESSION IN STABLY-TRANSFORMED LEPIDOPTERAN INSECT CELLS

2001 ◽  
Vol 37 (9) ◽  
pp. 564 ◽  
Author(s):  
SHUICHIRO TOMITA ◽  
YUKIE KAWAI ◽  
SOO DONG WOO ◽  
MANABU KAMIMURA ◽  
KIKUO IWABUCHI ◽  
...  
Keyword(s):  
Gene Expression ◽  
Insect Cells ◽  
Foreign Gene ◽  
Foreign Gene Expression ◽  
Lepidopteran Insect

scholarly journals Expression and Membrane Topology of Anopheles gambiae Odorant Receptors in Lepidopteran Insect Cells

PLoS ONE ◽  
2010 ◽  
Vol 5 (11) ◽  
pp. e15428 ◽  
Author(s):  
Panagiota Tsitoura ◽  
Evi Andronopoulou ◽  
Daniela Tsikou ◽  
Adamantia Agalou ◽  
Maria P. Papakonstantinou ◽  
...  
Keyword(s):  
Anopheles Gambiae ◽  
Insect Cells ◽  
Membrane Topology ◽  
Odorant Receptors ◽  
Lepidopteran Insect

scholarly journals Expression of a Toxoneuron nigriceps polydnavirus-encoded protein causes apoptosis-like programmed cell death in lepidopteran insect cells

2005 ◽  
Vol 86 (4) ◽  
pp. 963-971 ◽  
Author(s):  
Renée Lapointe ◽  
Rebecca Wilson ◽  
Lluïsa Vilaplana ◽  
David R. O'Reilly ◽  
Patrizia Falabella ◽  
...  
Keyword(s):  
Cell Death ◽  
Programmed Cell Death ◽  
Recombinant Virus ◽  
Insect Cells ◽  
Expression Patterns ◽  
Parental Virus ◽  
Tunel Staining ◽  
Lepidopteran Insect ◽  
High Five Cells ◽  
Toxoneuron Nigriceps

The polydnavirus Toxoneuron nigriceps bracovirus (TnBV) is an obligate symbiont associated with the braconid wasp T. nigriceps, a parasitoid of Heliothis virescens larvae. Previously, to identify polydnavirus genes that allow parasitization by altering the host immune and endocrine systems, expression patterns of TnBV genes from parasitized H. virescens larvae were analysed and cDNAs were obtained. To study the function of the protein from one such cDNA, TnBV1, overexpression of the protein was attempted by using the baculovirus Autographa californica multicapsid nucleopolyhedrovirus. Recovery of stable recombinant virus was unsuccessful, with the exception of recombinants with deletions/mutations within the TnBV1 gene. It was hypothesized that TnBV1 expression was cytotoxic to the Spodoptera frugiperda (Sf21) insect cells that were used to produce the recombinants. Therefore, the Bac-to-Bac system was used to create recombinant baculoviruses maintained in Escherichia coli expressing either TnBV1 (Ac-TnBV1) or an initiator-methionine mutant [Ac-TnBV1(ATG−)]. Microscopy revealed substantial cell death of Sf21 and High Five cells from 48 h post-infection with Ac-TnBV1, but not with the Ac-TnBV1(ATG−) recombinant virus. Ac-TnBV1-infected Sf21 cells, but not those with parental virus infection, showed an increased caspase-3-like protease activity, as well as increased terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling (TUNEL) for breaks in host genomic DNA. Although indicative of apoptosis, blebbing and apoptotic bodies were not observed in infected cells. Transiently expressing TnBV1 alone caused TUNEL staining in High Five cells. These data suggest that TnBV1 expression alone can induce apoptosis-like programmed cell death in two insect cell lines. Injection of Ac-TnBV1 budded virus, compared with parental virus, did not result in an alteration of virulence in H. virescens larvae.

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