RRC ID 36146
Author Imai S, Kusakabe T, Xu J, Li Z, Shirai S, Mon H, Morokuma D, Lee JM.
Title Roles of silkworm endoplasmic reticulum chaperones in the secretion of recombinant proteins expressed by baculovirus system.
Journal Mol Cell Biochem
Abstract Baculovirus expression vector system (BEVS) is widely used for production of recombinant eukaryotic proteins in insect larvae or cultured cells. BEVS has advantages over bacterial expression system in producing post-translationally modified secreted proteins. However, for some unknown reason, it is very difficult for insects to secrete sufficiently for certain proteins of interest. To understand the reasons why insect cells fail to secrete some kinds of recombinant proteins, we here employed three mammalian proteins as targets, EPO, HGF, and Wnt3A, with different secretion levels in BEVS and investigated their mRNA transcriptions from the viral genome, subcellular localizations, and interactions with silkworm ER chaperones. Moreover, we observed that no significantly influence on the secretion amounts of all three proteins when depleting or overexpressing most endogenous ER chaperone genes in cultured silkworm cells. However, among all detected ER chaperones, the depletion of BiP severely decreased the recombinant protein secretion in BEVS, indicating the possible central role of Bip in silkworm secretion pathway.
Volume 409(1-2)
Pages 255-62
Published 2015-11-1
DOI 10.1007/s11010-015-2529-5
PII 10.1007/s11010-015-2529-5
PMID 26264072
MeSH Animals Baculoviridae / genetics* Baculoviridae / metabolism Bombyx / genetics* Bombyx / metabolism Calnexin / genetics Calnexin / metabolism Calreticulin / genetics Calreticulin / metabolism Cats Cell Line Endoplasmic Reticulum / metabolism* Endoplasmic Reticulum Chaperone BiP Erythropoietin / genetics Erythropoietin / metabolism Genetic Vectors / genetics Heat-Shock Proteins / genetics Heat-Shock Proteins / metabolism Hepatocyte Growth Factor / genetics Hepatocyte Growth Factor / metabolism Humans Mice Molecular Chaperones / genetics Molecular Chaperones / metabolism* Protein Disulfide-Isomerases / genetics Protein Disulfide-Isomerases / metabolism RNA Interference RNA, Messenger / biosynthesis RNA, Messenger / genetics RNA, Small Interfering / genetics Recombinant Proteins / genetics Recombinant Proteins / metabolism* Wnt3A Protein / genetics Wnt3A Protein / metabolism
IF 2.795
Times Cited 6
Silkworms f38