Proton pump of clathrin-coated vesicles
Clathrin-coated vesicles were prepared from bovine brain catalyze ATP-driven proton translocation and a /sup 32/Pi-ATP exchange reaction. N-ethylmaleimide (NEM) at 1 mM and dicyclohexylcarbodiimide (DCCD) at 0.5 mM inhibit the pump completely, whereas neither vanadate, efrapeptin, NaN/sub 3/, nor mitochondrial ATPase inhibitor has an effect. The coated vesicle proton pump is characterized by ATP specificity. dATP, but no other nucleotide, can replace ATP as a substrate. The pump is electrogenic and the electrogenicity is neutralized by chloride or bromide serving as co-ions. ATP-driven proton translocation can be observed in the absence of chloride, provided that the membrane potential is collapsed by K/sup +/ moving out in the presence of valinomycin. Chloride transport can be observed independent of proton movements in the absence of ATP, provided that an inside positive membrane potential is generated by K/sup +/ influx in the presence of valinomycin. The proton-translocating ATPase of coated vesicles was solubilized with a nonionic detergent polyoxyethylene 9 lauryl ether, and purified about 700 fold to near homogeneity. During purification the enzymatic activity was lost. A purified brain phospholipid fraction restored the activity and was subsequently identified as phosphatidylserine.
- Research Organization:
- Cornell Univ., Ithaca, NY (USA)
- OSTI ID:
- 6023968
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
560301* -- Chemicals Metabolism & Toxicology-- Cells-- (-1987)
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ACID ANHYDRASES
ALKALI METAL COMPOUNDS
ANIMALS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
ATP
ATP-ASE
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
BODY
BRAIN
CATIONS
CATTLE
CENTRAL NERVOUS SYSTEM
CHARGED PARTICLES
CHLORIDES
CHLORINE COMPOUNDS
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
DRUGS
ENZYME ACTIVITY
ENZYMES
HALIDES
HALOGEN COMPOUNDS
HYDROLASES
IMIDES
IONS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MEMBRANE TRANSPORT
NERVOUS SYSTEM
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
OXYGEN COMPOUNDS
PHOSPHOHYDROLASES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
POTASSIUM COMPOUNDS
PURIFICATION
RADIOISOTOPES
RUMINANTS
TRACER TECHNIQUES
TRANSITION ELEMENT COMPOUNDS
VALINOMYCIN
VANADATES
VANADIUM COMPOUNDS
VERTEBRATES