RRC ID 4748
Author Shinya E, Owaki A, Norose Y, Sato S, Takahashi H.
Title Quick method of multimeric protein production for biologically active substances such as human GM-CSF (hGM-CSF).
Journal Biochem. Biophys. Res. Commun.
Abstract The C-terminal fragment of C4b-binding protein (C4BP)-based multimerizing system was applied to hGM-CSF to induce dendritic cells (DCs) from peripheral blood monocytes (PBMCs), to see whether the C4BP could stimulate immature DCs, since DCs, equipped with pattern recognition receptors such as toll-like receptors (TLRs), are hypersensitive to various immunologically active molecules like LPS. hGM-CSF gene was merged to the 3'-terminal region of the C4BPalpha-chain gene, and the transfected human 293FT cells produced sufficient amount of octameric hGM-CSF, which resulted in iDCs with the same phenotype and the same response to a TRL4 ligand, LPS and a TLR3 ligand, poly I:C, as those induced with authentic monomeric hGM-CSF. These results suggest that the C4BP-based multimerizing system could facilitate the design of self-associating multimeric recombinant proteins without stimulating iDCs, which might be seen with the other multimerizing systems such as that using Fc fragment of IgM.
Volume 386(1)
Pages 40-4
Published 2009-8-14
DOI 10.1016/j.bbrc.2009.05.125
PII S0006-291X(09)01089-4
PMID 19497303
MeSH Cell Line Complement C4b-Binding Protein / genetics Complement C4b-Binding Protein / metabolism Dendritic Cells / drug effects Dendritic Cells / metabolism Granulocyte-Macrophage Colony-Stimulating Factor / biosynthesis* Granulocyte-Macrophage Colony-Stimulating Factor / genetics Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology Humans Protein Biosynthesis* Protein Multimerization Recombinant Proteins / biosynthesis* Recombinant Proteins / genetics Recombinant Proteins / pharmacology
IF 2.559
Times Cited 1
WOS Category BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material pCM-hGM (RDB01687)