Use of inductively coupled plasma-mass spectrometry for the quantitation of the binding and uptake of colloidal gold-low-density lipoprotein conjugates by cultured cells.
The binding and endocytic uptake of low-density lipoprotein (LDL) particles by cells, transiently or permanently transfected with the human LDL receptor cDNA, was investigated, under different situations, using colloidal gold-LDL conjugates. The amount of gold associated with the various cells, which bind and internalize LDL to different extents, was estimated by inductively coupled plasma-MS. In all cases, the existing differences in LDL binding and uptake were clearly detectable with this procedure. We conclude, therefore, that inductively coupled plasma-MS provides an appropriate assay system for the rapid quantitation of these processes. This procedure also recognizes differences in LDL receptor expression in human lymphocytes and, therefore, it could be of value for the differential diagnosis of LDL receptor defects in familial hypercholesterolemia in various cell types. In addition, this easily performed methodology can also be applied to a variety of other problems requiring quantitation of colloidal gold associated with cells.
Martin de Llano
J J
JJ
Fundación Valenciana de Investigaciones Biomédicas, Instituto de Investigaciones Citológicas, Amadeo de Saboya 4, Valencia, 46010, Spain.
Andreu
E J
EJ
Knecht
E
E
D000818:Animals; D001693:Biological Transport, Active; D016466:CHO Cells; D019556:COS Cells; D002417:Cattle; D002478:Cells, Cultured; D006224:Cricetinae; D004705:Endocytosis; D017979:Gold Colloid / Q000378:metabolism*; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D008854:Microscopy, Electron; D011973:Receptors, LDL / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D017979:Gold Colloid; D008077:Lipoproteins, LDL; D011973:Receptors, LDL