| RRC ID |
42636
|
| 著者 |
Yura H, Ishihara M, Kanatani Y, Takase B, Hattori H, Suzuki S, Kawakami M, Matsui T.
|
| タイトル |
Interaction study between synthetic glycoconjugate ligands and endocytic receptors using flow cytometry.
|
| ジャーナル |
J Biochem
|
| Abstract |
Flow cytometric analysis of synthetic galactosyl polymers, asialofetuin and LDL derivatives labeled with FITC (Fluorescein Isothiocyanate) was carried out to determine the phenotypes of endocytic receptors, such as asialoglycoprotein (ASPG) and the LDL receptor, on various types of cells. When FITC-labeled galactosyl polystyrene (GalCPS), being a synthetic ligand of ASPG, was applied to rat hepatocytes and human cancer cells (Hep G2 and Chang Liver), surface fluorescence intensities varied according to receptor expression on the cells. The fluorescence intensity originates from the calcium-dependent binding of the FITC-labeled GalCPS. Although unaltered by pre-treatment with glucosyl polystyrene (GluCPS), fetuin and LDL, the fluorescence intensity was suppressed by pre-treatment with (non-labeled) GalCPS and asialofetuin. Flow cytometry allowed us to demonstrate that the calcium-dependent binding of FITC-labeled LDL (prepared from rabbits) upon the addition of 17alpha-ethinyl estradiol enhances LDL receptor expression, and the expression is suppressed upon the addition of a monoclonal antibody to the LDL receptor. The binding efficiency based on the combination of FITC-labeled ligands suggests a possible application for the classification of cell types and conditions corresponding to endocytic receptor expression without the need for immuno-active antibodies or radiolabeled substances. Furthermore, the synthetic glycoconjugate (GalCPS) is shown to be a sensitive and useful marker for classification based on cell phenotype using flow cytometry.
|
| 巻・号 |
139(4)
|
| ページ |
637-43
|
| 公開日 |
2006-4-1
|
| DOI |
10.1093/jb/mvj077
|
| PII |
139/4/637
|
| PMID |
16672263
|
| MeSH |
Animals
Asialoglycoproteins / chemistry
Asialoglycoproteins / metabolism
Binding, Competitive
Cell Line, Tumor
Cells, Cultured
Female
Fetuins
Flow Cytometry / methods*
Fluorescein-5-isothiocyanate / chemistry
Fluorescent Antibody Technique / methods
Glycoconjugates / chemical synthesis
Glycoconjugates / chemistry
Glycoconjugates / metabolism*
Hepatocytes / cytology
Hepatocytes / metabolism
Humans
Lipoproteins, LDL / chemistry
Lipoproteins, LDL / metabolism
Molecular Structure
Protein Binding
Rabbits
Rats
Rats, Sprague-Dawley
Receptors, Cell Surface / metabolism*
Receptors, LDL / metabolism
Receptors, Lipoprotein / metabolism
alpha-Fetoproteins / chemistry
alpha-Fetoproteins / metabolism
|
| IF |
2.476
|
| 引用数 |
5
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
Hep G2 |
