Reference - Detail
RRC ID | 42570 |
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Author | Matsunaga T, Kose E, Yasuda S, Ise H, Ikeda U, Ohmori S. |
Title | Determination of p-glycoprotein ATPase activity using luciferase. |
Journal | Biol Pharm Bull |
Abstract |
We investigated whether P-glycoprotein (P-gp) ATPase activity of Caco-2 cell membranes could be estimated by measuring consumption of ATP using luciferin-luciferase reaction, and whether the results would be useful for assessment of the interactions between P-gp and drugs. The vanadate-sensitive ATPase activity of Caco-2 cell membranes was measured rapidly with high sensitivity using luciferin-luciferase reaction. Cyclosporin A, verapamil, digoxin and quinidine stimulated the ATPase activity concentration-dependently with Km values of 5.3, 0.9, 1.2 and 4.1 microM, respectively. These values except for digoxin were comparable with previous reports. The ATPase activity and P-gp mRNA expression in Caco-2 cells were induced by all-trans-retinoic acid, digoxin and levothyroxine, but not dexamethasone or rifampicin. This method was useful to assess interactions with P-gp and drugs, and was used to elucidate the mechanisms of interaction of levothyroxine and digoxin. |
Volume | 29(3) |
Pages | 560-4 |
Published | 2006-3-1 |
DOI | 10.1248/bpb.29.560 |
PII | JST.JSTAGE/bpb/29.560 |
PMID | 16508168 |
MeSH | ATP Binding Cassette Transporter, Subfamily B, Member 1 / analysis* ATP Binding Cassette Transporter, Subfamily B, Member 1 / metabolism Adenosine Triphosphatases / analysis* Adenosine Triphosphatases / metabolism Adenosine Triphosphate / metabolism Caco-2 Cells Cell Membrane / drug effects Cell Membrane / metabolism Dexamethasone / pharmacology Digoxin / pharmacology Glucocorticoids / pharmacology Humans Kinetics Luciferases* RNA, Messenger / biosynthesis RNA, Messenger / isolation & purification Rifampin / pharmacology Thyroxine / pharmacology |
IF | 1.863 |
Times Cited | 12 |
WOS Category | PHARMACOLOGY & PHARMACY |
Resource | |
Human and Animal Cells | CACO-2(RCB0988) |