Reference - Detail
| 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 |
| Resource | |
| Human and Animal Cells | COS-1(RCB0143) |