Rapid preparation of a plasma membrane fraction from adipocytes and muscle cells: application to detection of translocated glucose transporter 4 on the plasma membrane.
The aim of this study was to establish a rapid preparation of plasma membrane from adipocytes and muscle cells to detect translocated glucose transporter (GLUT) 4. A plasma membrane fraction was prepared by sequential centrifugation with buffer containing detergents, and its purity was estimated by detecting insulin receptor beta-subunit (IRbeta). After insulin stimulus, GLUT4 translocation was observed in 3T3-L1 adipocytes and L6 myotubes. It was found that IRbeta and GLUT4 levels on the plasma membrane decreased in adipose and muscle with intake of a 29% lard diet for 14 weeks. Hence, this method should be useful for rapid preparation of the plasma membrane fraction.
Nishiumi
Shin
S
Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Hyogo 657-8501, Japan.
Ashida
Hitoshi
H
D017667:Adipocytes / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D045304:Cytochromes c / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D005223:Fats / Q000494:pharmacology; D051275:Glucose Transporter Type 4 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D032342:Muscle Cells / Q000378:metabolism*; D014176:Protein Biosynthesis; D021381:Protein Transport; D051381:Rats; D011972:Receptor, Insulin / Q000378:metabolism; D013347:Subcellular Fractions; D013997:Time Factors