PMID: 6170323Sep 1, 1981Paper

Binding, surface mobility, internalization, and degradation of rhodamine-labeled alpha 2-macroglobulin

Biochemistry
F R MaxfieldI Pastan

Abstract

We have used quantitative fluorescence methods to examine the fate of rhodamine-labeled alpha 2-macroglobulin (R-alpha 2 M) after binding to cell-surface receptors on NRK and Swiss 3T3 cells. From measurements of fluorescence intensities in NRK cells fixed after incubation with R-alpha 2M, we found that uptake was saturable and that half-maximal uptake occurred at 130 nM R-alpha 2M. Fluorescence measurements on cell extracts of NRK and Swiss 3T3 cells also showed a half-maximal uptake of R-alpha 2M near 130 nM. We estimate that NRK cells can take up 10(6) molecules of R-alpha 2M per hour via receptor-mediated endocytosis. The mobility of alpha 2-macroglobulin receptors on the surface of Swiss 3T3 cells was measured by using fluorescence photobleaching recovery. The two-dimensional effective diffusion coefficient of R-alpha 2M receptors was approximately 8 X 10(-10) cm2 s-1, a value close to that previously obtained for insulin and epidermal growth factor receptors. Degradation of R-alpha 2M by the cells was followed by using the loss of fluorescence from the 185000-dalton band in sodium dodecyl sulfate--polyacrylamide gels. Rhodamine fluorescence was detected in the gels by using a microscope fluorescence spectrophotometer. NRK...Continue Reading

Citations

Jan 1, 1984·Molecular and Cellular Biochemistry·P J Davies, M P Murtaugh
Feb 1, 1983·The Histochemical Journal·W StauberF Gauthier
Jan 1, 1983·The Journal of Membrane Biology·D Axelrod
Mar 31, 1983·Biochimica Et Biophysica Acta·J GliemannL Sottrup-Jensen
Jan 1, 1984·Collagen and Related Research·W Borth
Aug 5, 2009·Journal of Leukocyte Biology·Edith V BowersSalvatore V Pizzo
Jul 1, 1984·Experimental Cell Research·S SmitB Leijnse
Jan 1, 1983·Annals of the New York Academy of Sciences·B TyckoF R Maxfield
Nov 22, 2005·Molecular and Cellular Endocrinology·Chandrakant TayadeB A Croy
Aug 2, 2005·Journal of Biotechnology·Moltira TonganuntKenichi Fujise
Nov 20, 1997·The Journal of Biological Chemistry·D M ParesceF R Maxfield
Sep 10, 1994·Annals of the New York Academy of Sciences·C T ChuS V Pizzo
May 1, 1983·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·C J Steer, R D Klausner
Nov 12, 1985·Biochimica Et Biophysica Acta·O Saksela

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