RRC ID |
534
|
著者 |
Abo Y, Hagiya A, Naganuma T, Tohkairin Y, Shiomi K, Kajiura Z, Hachimori A, Uchiumi T, Nakagaki M.
|
タイトル |
Baculovirus-mediated expression and isolation of human ribosomal phosphoprotein P0 carrying a GST-tag in a functional state.
|
ジャーナル |
Biochem Biophys Res Commun
|
Abstract |
We constructed an overexpression system for human ribosomal phosphoprotein P0, together with P1 and P2, which is crucially important for translation. Genes for these proteins, fused with the glutathione S-transferase (GST)-tag at the N-terminus, were inserted into baculovirus and introduced to insect cells. The fusion proteins, but not the proteins without the tag, were efficiently expressed into cells as soluble forms. The fusion protein GST.P0 as well as GST.P1/GST.P2 was phosphorylated in cells as detected by incorporation of (32)P and reactivity with monoclonal anti-phosphoserine antibody. GST.P0 expressed in insect cells, but not the protein obtained in Escherichia coli, had the ability to form a complex with P1 and P2 proteins and to bind to 28S rRNA. Moreover, the GST.P0-P1-P2 complex participated in high eEF-2-dependent GTPase activity. Baculovirus expression systems appear to provide recombinant human P0 samples that can be used for studies on the structure and function.
|
巻・号 |
322(3)
|
ページ |
814-9
|
公開日 |
2004-9-24
|
DOI |
10.1016/j.bbrc.2004.07.196
|
PII |
S0006-291X(04)01728-0
|
PMID |
15336536
|
MeSH |
Animals
Baculoviridae / genetics*
Cell Line
Gene Expression Regulation, Viral / genetics
Genetic Vectors
Glutathione Transferase / genetics
Humans
Phosphoproteins / genetics*
Phosphoproteins / isolation & purification
Phosphoproteins / metabolism
Recombinant Fusion Proteins / isolation & purification
Recombinant Fusion Proteins / metabolism
Ribosomal Proteins / genetics*
Ribosomal Proteins / isolation & purification
Ribosomal Proteins / metabolism
Spodoptera
Transfection
|
IF |
2.985
|
引用数 |
2
|
WOS 分野
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
カイコ |
|