Functional reconstitution of yeast mitochondrial ADP/ATP carrier by removing detergent with Amberlite treatment. 1991

K Narayanareddy
Institute for physical Biochemistry, University of Munich, Germany.

The ADP/ATP carrier (AAC) from yeast mitochondria has been reconstituted in phospholipid vesicles essentially according to the procedure described for the reconstitution of AAC from bovine heart mitochondria (Krämer and Heberger (1986) Biochim. Biophys. Acta, 863, 289-296). Liposomes were prepared from the mixed micelles of dodecyl octaoxyethylene ether (C12E8)-solubilized protein and egg yolk phosphatidylcholine by removing the detergent with Amberlite treatment. The micelles were treated with Amberlite either by repeatedly passing through small columns filled with Amberlite XAD-2 beads or by stepwise addition of Amberlite beads to the micelles. All the important variables of the reconstitution components were kept at optimal level and the liposomes obtained by both the methods of Amberlite treatment were analysed for (3H)CAT binding, orientation of AAC and nucleotide exchange activity. Reconstituted AAC showed 80% right side out orientation in the liposomes prepared by either procedure. Lipsomes prepared by the Amberlite column procedure exhibited higher CAT binding but lower ADP exchange activity. Liposome preparation by the stepwise addition of Amberlite is suggested to be the method of choice for functional reconstitution of yeast AAC in view of the higher nucleotide transport activity associated with the liposomes prepared by this method.

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
D008928 Mitochondria Semiautonomous, self-reproducing organelles that occur in the cytoplasm of all cells of most, but not all, eukaryotes. Each mitochondrion is surrounded by a double limiting membrane. The inner membrane is highly invaginated, and its projections are called cristae. Mitochondria are the sites of the reactions of oxidative phosphorylation, which result in the formation of ATP. They contain distinctive RIBOSOMES, transfer RNAs (RNA, TRANSFER); AMINO ACYL T RNA SYNTHETASES; and elongation and termination factors. Mitochondria depend upon genes within the nucleus of the cells in which they reside for many essential messenger RNAs (RNA, MESSENGER). Mitochondria are believed to have arisen from aerobic bacteria that established a symbiotic relationship with primitive protoeukaryotes. (King & Stansfield, A Dictionary of Genetics, 4th ed) Mitochondrial Contraction,Mitochondrion,Contraction, Mitochondrial,Contractions, Mitochondrial,Mitochondrial Contractions
D012117 Resins, Synthetic Polymers of high molecular weight which at some stage are capable of being molded and then harden to form useful components. Dental Resins,Dental Resin,Resin, Dental,Resin, Synthetic,Resins, Dental,Synthetic Resin,Synthetic Resins
D003902 Detergents Purifying or cleansing agents, usually salts of long-chain aliphatic bases or acids, that exert cleansing (oil-dissolving) and antimicrobial effects through a surface action that depends on possessing both hydrophilic and hydrophobic properties. Cleansing Agents,Detergent Pods,Laundry Detergent Pods,Laundry Pods,Syndet,Synthetic Detergent,Agent, Cleansing,Agents, Cleansing,Cleansing Agent,Detergent,Detergent Pod,Detergent Pod, Laundry,Detergent Pods, Laundry,Detergent, Synthetic,Detergents, Synthetic,Laundry Detergent Pod,Laundry Pod,Pod, Detergent,Pod, Laundry,Pod, Laundry Detergent,Pods, Detergent,Pods, Laundry,Pods, Laundry Detergent,Synthetic Detergents
D000226 Mitochondrial ADP, ATP Translocases A class of nucleotide translocases found abundantly in mitochondria that function as integral components of the inner mitochondrial membrane. They facilitate the exchange of ADP and ATP between the cytosol and the mitochondria, thereby linking the subcellular compartments of ATP production to those of ATP utilization. ADP,ATP Carrier,ADP,ATP Translocator Protein,Adenine Nucleotide Translocase,ADP Translocase,ATP Translocase,ATP,ADP-Carrier,ATP-ADP Translocase,Adenine Nucleotide Carrier (Mitochondrial),Mitochondrial ADP-ATP Carriers,ADP-ATP Carriers, Mitochondrial,Mitochondrial ADP ATP Carriers
D001278 Atractyloside A glycoside of a kaurene type diterpene that is found in some plants including Atractylis gummifera (ATRACTYLIS); COFFEE; XANTHIUM, and CALLILEPIS. Toxicity is due to inhibition of ADENINE NUCLEOTIDE TRANSLOCASE. Atractylic Acid,Atractylate,Acid, Atractylic
D012441 Saccharomyces cerevisiae A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement. Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker

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