Delivery of liposome-sequestered hydrophobic proteins to lysosomes of normal and Batten disease cells.
We have developed a method to deliver hydrophobic proteins such as ATP synthase subunit c and ubiquitin to lysosomes of PMN (polymorphonucleocytes) and fibroblasts. ATP synthase subunit c is stored in the lysosomes of various tissues in late infantile and juvenile forms of neuronal ceriod lipofuscinosis, also called Batten disease (BD). Whether this protein storage is due to an abbreviation in protein or in the lysosomal hydrolases of BD is still not clear. We have sequestered this protein and ubiquitin in the lipid membrane of liposomes. The liposomes coated with autologous heat-aggregated IgG or apolipoprotein E when presented to the PMN and fibroblasts, respectively, accumulated in the lysosomes. Both normal and BD PMN degraded 125I-ubiquitin; the rate of degradation was, however, slower by Batten PMN. These studies indicate that a hydrophobic molecule such as subunit c can be delivered to PMN and fibroblasts, and the sequestered proteins are accessible to lysosomal hydrolases. Therefore, this technique can be used to study the metabolism of highly hydrophobic proteins by lysosomes, especially the biochemical mechanism(s) of subunit c storage in BD.
Ansari
N H
NH
Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston 77555-0647, USA.
He
Q
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Cook
J D
JD
Wen
J
J
Srivastava
S K
SK
D001057:Apolipoproteins E; D002451:Cell Compartmentation / Q000502:physiology; D002478:Cells, Cultured / Q000201:enzymology / Q000648:ultrastructure; D002850:Chromatography, Gel; D003911:Dextrans; D005347:Fibroblasts / Q000201:enzymology / Q000648:ultrastructure; D005782:Gels; D006801:Humans; D006867:Hydrolases / Q000378:metabolism; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D008081:Liposomes / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism*; D008854:Microscopy, Electron; D009472:Neuronal Ceroid-Lipofuscinoses / Q000378:metabolism*; D009504:Neutrophils / Q000201:enzymology / Q000648:ultrastructure; D011233:Precipitin Tests; D006180:Proton-Translocating ATPases / Q000378:metabolism; D012867:Skin / Q000166:cytology; D014452:Ubiquitins / Q000378:metabolism
D001057:Apolipoproteins E; D003911:Dextrans; D005782:Gels; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D008081:Liposomes; D014452:Ubiquitins; C025614:sephadex; D006867:Hydrolases; D006180:Proton-Translocating ATPases