PMID: 8948454Jun 15, 1995Paper

Identification of a lysosomal protein causing lipid transfer, using a fluorescence assay designed to monitor membrane fusion between rat liver endosomes and lysosomes

The Biochemical Journal
T KuwanaJ Paul Luzio

Abstract

In the present and previous studies [Mullock, Perez, Kuwana, Gray and Luzio (1994) J. Cell Biol. 126, 1173-1182], we have attempted to investigate endosome-lysosome fusion using an assay based on the dilution of the self-quenching fluorescent lipid probe octadecylrhodamine. Although some characteristics of fluorescence dequenching were consistent with those observed in other cell-free assays, we have now demonstrated that increased fluorescence was due to leakage of an intralysosomal lipid-transfer protein. This protein was purified and found to be a 22 kDa molecule with sequence, immunological and functional characteristics strongly suggesting that it is the rat homologue of human GM2-activator protein. Both the 22 kDa protein and recombinant human GM2-activator protein caused fluorescence dequenching either when mixed with octadecylrhodamine-loaded endosomes and lysosomal membranes or in a liposome system. The data were consistent with GM2-activator protein acting as an octadecylrhodamine-transfer protein. Antibodies to the 22 kDa protein added to cell-free endosome-lysosome content-mixing assays had no effect, although they could inhibit fluorescence dequenching caused by the protein. Thus this protein is not required in any...Continue Reading

Citations

Feb 1, 2011·Journal of Drug Targeting·Alexander KoshkaryevVladimir P Torchilin
Sep 21, 2010·Chemistry and Physics of Lipids·Irene TrikashValeriy Lishko
May 4, 2002·Biochimica Et Biophysica Acta·Isabel Nunes-CorreiaMaria C Pedroso de Lima
Sep 15, 2011·Bioconjugate Chemistry·Igor MeerovichVladimir P Torchilin
Feb 1, 1997·Mammalian Genome : Official Journal of the International Mammalian Genome Society·C BertoniT Beccari
May 18, 1999·Biochemical and Biophysical Research Communications·B RigatD Mahuran
May 23, 1997·Biochimica Et Biophysica Acta·N Smiljanic-GeorgijevD J Mahuran
Mar 12, 1999·Journal of the American Society of Nephrology : JASN·T M MundelP Mundel

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