ATP-dependent transport of vinblastine in vesicles from human multidrug-resistant cells
- National Institutes of Health, Bethesda, MD (USA)
Resistance of human cancer cells to multiple cytotoxic hydrophobic agents (multidrug resistance) is due to overexpression of the MDR1 gene, whose product is the plasma membrane P-glycoprotein. Plasma membrane vesicles partially purified from multidrug-resistant human KB carcinoma cells, but not from drug-sensitive cells, accumulate ({sup 3}H)vinblastine in an ATP-dependent manner. This transport is osmotically sensitive, with an apparent K{sub m} of 38 {mu}M for ATP and of {approx} 2 {mu}M for vinblastine. The nonhydrolyzable analog adenosine 5{prime}-({beta},{gamma}-imido)triphosphate does not substitute for ATP but is a competitive inhibitor of ATP for the transport process. Vanadate, and ATPase inhibitor, is a potent noncompetitive inhibitor of transport. These results indicate that hydrolysis of ATP is probably required for active transport vinblastine. Several other drugs to which multidrug-resistant cell lines are resistant inhibit transport, with relative potencies as follows: vincristine > actinomycin D > daunomycin > colchicine = puromycin. Verapamil and quinidine, which reverse the multidrug-resistance phenotype, are good inhibitors of the transport process. These results confirm that multidrug-resistant cells express an energy-dependent plasma membrane transporter for hydrophobic drugs, and establish a system for the detailed biochemical analysis of this transport process.
- OSTI ID:
- 5517837
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 85:10; ISSN 0027-8424
- Country of Publication:
- United States
- Language:
- English
Similar Records
Molecular basis of preferential resistance to colchicine in multidrug-resistant human cells conferred by Gly-185 yields Val-185 substitution in P-glycoprotein
Functional expression of murine multidrug resistance in Xenopus laevis oocytes
Related Subjects
ATP-ASE
BIOCHEMICAL REACTION KINETICS
TRITIUM COMPOUNDS
MEMBRANE TRANSPORT
TUMOR CELLS
SENSITIVITY
VINBLASTINE
ATP
ENZYME INHIBITORS
GLUCOPROTEINS
MAN
VANADATES
ACID ANHYDRASES
ALKALOIDS
ANIMAL CELLS
ANIMALS
ANTIMITOTIC DRUGS
AROMATICS
AZAARENES
AZOLES
CARBOHYDRATES
DRUGS
ENZYMES
GLYCOPROTEINS
HETEROCYCLIC COMPOUNDS
HYDROGEN COMPOUNDS
HYDROLASES
INDOLES
KINETICS
MAMMALS
NUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
OXYGEN COMPOUNDS
PHOSPHOHYDROLASES
PRIMATES
PROTEINS
PYRROLES
REACTION KINETICS
SACCHARIDES
TRANSITION ELEMENT COMPOUNDS
VANADIUM COMPOUNDS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques