Enzymatic lysis of sulfated glycosaminoglycans reduces the electrophoretic mobility of vascular endothelial cells

Membrane Biochemistry
F F VargasU S Ryan

Abstract

The main purpose of this work was to identify the macromolecules carrying the surface charge of endothelial cells. This was done by measuring changes in cell electrophoretic mobility caused by enzymatic removal of glycocalyx components. Endothelial cells were removed from the bovine pulmonary artery using nonenzymatic procedures, plated, and identified by immunocytochemical methods and electron microscopy. Cultured cells were suspended in saline and placed in the lumen of a capillary in a Rank Brothers electrophoresis instrument. Voltage was applied between the ends of the capillary, and the velocity acquired by the cells was measured with a microscope. Preincubating the cells in protein-free saline for 1 h reduced the mobility by 25%. This reflects the loss of proteoheparan sulfate from the cell surface. Cell mobility was totally suppressed by exposing the entire cell surface to chondroitin sulfate lyase, but it was only slightly diminished when the enzyme was applied only to the cell side facing the culture medium. A partial decrease in mobility was obtained after enzymatic removal of either heparin, heparan sulfate, or collagen. The results indicate that sulfated glycosaminoglycans are the main carriers of the surface change...Continue Reading

References

Jan 1, 1989·Membrane Biochemistry·F F VargasM De Jesus
Jan 1, 1989·Annals of the New York Academy of Sciences·J E SilbertC K Silbert
Jan 1, 1985·The Journal of Membrane Biology·S L Mironov, E V Dolgaya
Dec 27, 1983·Biochimica Et Biophysica Acta·D T WoodleyH K Kleinman
Jan 1, 1984·Annual Review of Biochemistry·M HöökS Johansson

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Citations

Mar 19, 2008·Journal of Cellular Biochemistry·Kristin L ReigleJames D San Antonio
Mar 14, 2000·The Journal of Biological Chemistry·A VerrecchioJ D San Antonio
Feb 8, 2008·Journal of Biomedical Materials Research. Part a·Dong-Hyun KimYong-Keun Lee
Jun 1, 1997·Electrophoresis·G G SlivinskyD R Morrison

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