RRC ID 76396
Author Maeda Y, Ueda H, Kazami J, Kawano G, Suzuki E, Nagamune T.
Title Engineering of functional chimeric protein G-Vargula luciferase.
Journal Anal Biochem
Abstract Luciferase of Vargula hilgendorfli is infinitely stable at room temperature in dried state, and its light-emitting reaction is very simple. These unique characteristics of Vargula luciferase have prompted us to engineer chimeric protein, the other moiety chosen for conjugation being streptococcal protein G. A single domain of protein G which binds to IgG of a wide range of species was fused at the N-terminal region of Vargula luciferase. Unexpectedly, we found that the chimeric protein expressed in mammalian COS-1 cells had no IgG-binding ability, probably due to some sort of interaction between the two moieties or some conformational preferences of the IgG-binding domain of protein G when fused to Vargula luciferase. Here we report how we regained the IgG binding of protein G, by the intervention of three alpha-helices of protein A between protein G and luciferase. To our knowledge, the new chimeric protein provides the first reported model of this kind.
Volume 249(2)
Pages 147-52
Published 1997-7-1
DOI 10.1006/abio.1997.2181
PII S0003-2697(97)92181-3
PMID 9212866
MeSH Animals COS Cells Crustacea / enzymology* DNA, Recombinant Genetic Vectors Immunoglobulin Fc Fragments / metabolism Immunoglobulin G / metabolism Luciferases / biosynthesis Luciferases / genetics* Luciferases / metabolism Nerve Tissue Proteins / biosynthesis Nerve Tissue Proteins / genetics* Nerve Tissue Proteins / metabolism Protein Binding Protein Engineering* / methods Recombinant Fusion Proteins / biosynthesis Recombinant Fusion Proteins / genetics* Recombinant Fusion Proteins / metabolism Staphylococcal Protein A / genetics Staphylococcal Protein A / metabolism Streptococcus Transfection
IF 2.877
Human and Animal Cells COS-1(RCB0143)