Reconstitution of lysosomal sulfate transport in proteoliposomes. 1995

P J Koetters, and H F Chou, and A J Jonas
Department of Pediatrics, Harbor-UCLA Medical Center, Torrance 90502, USA.

As part of a strategy to purify the lysosomal sulfate transporter, we developed a method for reconstitution of transport in artificial membrane vesicles. Lysosomal membranes were prepared from Percoll density gradient purified rat liver lysosomes and membrane proteins were solubilized using the non-ionic detergent, Triton X-100. The solubilized proteins were mixed with liposomes prepared by sonication of egg yolk lecithin and the detergent was removed by passage of the mixture over Bio-beads XAD2. The resulting proteoliposomes exhibited saturable sulfate transport with characteristics that were very similar to those observed in lysosomal membranes. Transport in proteoliposomes had a Km of 155 microM, exhibited pH dependence and was sensitive to inhibition by DIDS. Reconstitution of transport in proteoliposomes may be useful as an assay for purification of the lysosomal sulfate carrier.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008081 Liposomes Artificial, single or multilaminar vesicles (made from lecithins or other lipids) that are used for the delivery of a variety of biological molecules or molecular complexes to cells, for example, drug delivery and gene transfer. They are also used to study membranes and membrane proteins. Niosomes,Transferosomes,Ultradeformable Liposomes,Liposomes, Ultra-deformable,Liposome,Liposome, Ultra-deformable,Liposome, Ultradeformable,Liposomes, Ultra deformable,Liposomes, Ultradeformable,Niosome,Transferosome,Ultra-deformable Liposome,Ultra-deformable Liposomes,Ultradeformable Liposome
D008099 Liver A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances. Livers
D008247 Lysosomes A class of morphologically heterogeneous cytoplasmic particles in animal and plant tissues characterized by their content of hydrolytic enzymes and the structure-linked latency of these enzymes. The intracellular functions of lysosomes depend on their lytic potential. The single unit membrane of the lysosome acts as a barrier between the enzymes enclosed in the lysosome and the external substrate. The activity of the enzymes contained in lysosomes is limited or nil unless the vesicle in which they are enclosed is ruptured or undergoes MEMBRANE FUSION. (From Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed). Autolysosome,Autolysosomes,Lysosome
D002499 Centrifugation, Density Gradient Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Centrifugations, Density Gradient,Density Gradient Centrifugation,Density Gradient Centrifugations,Gradient Centrifugation, Density,Gradient Centrifugations, Density
D005260 Female Females
D006863 Hydrogen-Ion Concentration The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001692 Biological Transport The movement of materials (including biochemical substances and drugs) through a biological system at the cellular level. The transport can be across cell membranes and epithelial layers. It also can occur within intracellular compartments and extracellular compartments. Transport, Biological,Biologic Transport,Transport, Biologic
D013431 Sulfates Inorganic salts of sulfuric acid. Sulfate,Sulfates, Inorganic,Inorganic Sulfates

Related Publications

P J Koetters, and H F Chou, and A J Jonas
July 1991, FEBS letters,
P J Koetters, and H F Chou, and A J Jonas
July 1992, Proceedings of the National Academy of Sciences of the United States of America,
P J Koetters, and H F Chou, and A J Jonas
June 2002, Biological chemistry,
P J Koetters, and H F Chou, and A J Jonas
February 1992, Biochimica et biophysica acta,
P J Koetters, and H F Chou, and A J Jonas
May 1995, The Journal of biological chemistry,
P J Koetters, and H F Chou, and A J Jonas
September 1994, The Journal of biological chemistry,
P J Koetters, and H F Chou, and A J Jonas
October 1988, The Journal of biological chemistry,
P J Koetters, and H F Chou, and A J Jonas
November 1993, The Journal of biological chemistry,
P J Koetters, and H F Chou, and A J Jonas
January 2004, Scandinavian journal of clinical and laboratory investigation,
Copied contents to your clipboard!