Nucleotide triphosphatase activity of the N-terminal nucleotide-binding domains of the multidrug resistance proteins P-glycoprotein and MRP1

Denise M. Wilkes; Changsen Wang; Patricia C. Aristimuo; Ariel F. Castro; Guillermo A. Altenberg

doi:10.1016/S0006-291X(02)00878-1

Nucleotide triphosphatase activity of the N-terminal nucleotide-binding domains of the multidrug resistance proteins P-glycoprotein and MRP1

Denise M. Wilkes, Changsen Wang, Patricia C. Aristimuo, Ariel F. Castro, Guillermo A. Altenberg

Anesthesiology

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

Abstract

The multidrug resistance proteins P-glycoprotein (Pgp) and MRP1 are drug-efflux pumps. In this study, we compared the nucleotide triphosphatase activities of the isolated N-terminal nucleotide binding domains (NBD1) of Pgp and MRP1, and explored the potential role of the phosphorylation target domain of Pgp on the regulation of Pgp NBD1 ATPase activity. We found that: (1) the NBD1s of Pgp and MRP1 have ATPase and GTPase activities, (2) the K_ms of Pgp NBD1 for ATP and GTP hydrolysis are identical, while the K_m of MRP1 NBD1 for ATP is lower than that for GTP, and (3) phosphorylation of MLD by PKA or PKC produces a marginal increase of V_max for ATP hydrolysis, without affecting the affinity for ATP. These results show efficient GTP hydrolysis by the NBD1s of Pgp and MRP1, and a minor role of phosphorylation in the control of Pgp NBD1 ATPase activity.

Original language	English (US)
Pages (from-to)	388-394
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	296
Issue number	2
DOIs	https://doi.org/10.1016/S0006-291X(02)00878-1
State	Published - 2002

Keywords

ABC proteins
CFTR
Expression
Maltose-binding protein
Multidrug resistance
Nucleotide-binding fold
Phosphorylation
Purification
R domain

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/S0006-291X(02)00878-1

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@article{6f66eb2ef7ed4db6a83369e348a84837,

title = "Nucleotide triphosphatase activity of the N-terminal nucleotide-binding domains of the multidrug resistance proteins P-glycoprotein and MRP1",

abstract = "The multidrug resistance proteins P-glycoprotein (Pgp) and MRP1 are drug-efflux pumps. In this study, we compared the nucleotide triphosphatase activities of the isolated N-terminal nucleotide binding domains (NBD1) of Pgp and MRP1, and explored the potential role of the phosphorylation target domain of Pgp on the regulation of Pgp NBD1 ATPase activity. We found that: (1) the NBD1s of Pgp and MRP1 have ATPase and GTPase activities, (2) the Kms of Pgp NBD1 for ATP and GTP hydrolysis are identical, while the Km of MRP1 NBD1 for ATP is lower than that for GTP, and (3) phosphorylation of MLD by PKA or PKC produces a marginal increase of Vmax for ATP hydrolysis, without affecting the affinity for ATP. These results show efficient GTP hydrolysis by the NBD1s of Pgp and MRP1, and a minor role of phosphorylation in the control of Pgp NBD1 ATPase activity.",

keywords = "ABC proteins, CFTR, Expression, Maltose-binding protein, Multidrug resistance, Nucleotide-binding fold, Phosphorylation, Purification, R domain",

author = "Wilkes, {Denise M.} and Changsen Wang and Aristimuo, {Patricia C.} and Castro, {Ariel F.} and Altenberg, {Guillermo A.}",

note = "Funding Information: This work was supported in part by grants from the National Cancer Institute and the American Heart Association, Texas Affiliate, Inc. We thank Dr. L. Reuss for comments on a preliminary version of the manuscript. We also wish to thank Dr. N. Greenfield (Circular Dichroism Facility, University of Medicine and Dentistry of New Jersey) for the circular dichroism spectroscopy, Dr. P. Borst and M. Kool (The Netherlands Cancer Institute) for providing us with the MRP1 cDNA, and J.S. Smith (Protein Chemistry Laboratory at the University of Texas medical Branch) for the peptide sequencing. We also thank C. Magliolo for technical assistance.",

year = "2002",

doi = "10.1016/S0006-291X(02)00878-1",

language = "English (US)",

volume = "296",

pages = "388--394",

journal = "Biochemical and Biophysical Research Communications",

issn = "0006-291X",

publisher = "Elsevier B.V.",

number = "2",

}

TY - JOUR

T1 - Nucleotide triphosphatase activity of the N-terminal nucleotide-binding domains of the multidrug resistance proteins P-glycoprotein and MRP1

AU - Wilkes, Denise M.

AU - Wang, Changsen

AU - Aristimuo, Patricia C.

AU - Castro, Ariel F.

AU - Altenberg, Guillermo A.

N1 - Funding Information: This work was supported in part by grants from the National Cancer Institute and the American Heart Association, Texas Affiliate, Inc. We thank Dr. L. Reuss for comments on a preliminary version of the manuscript. We also wish to thank Dr. N. Greenfield (Circular Dichroism Facility, University of Medicine and Dentistry of New Jersey) for the circular dichroism spectroscopy, Dr. P. Borst and M. Kool (The Netherlands Cancer Institute) for providing us with the MRP1 cDNA, and J.S. Smith (Protein Chemistry Laboratory at the University of Texas medical Branch) for the peptide sequencing. We also thank C. Magliolo for technical assistance.

PY - 2002

Y1 - 2002

N2 - The multidrug resistance proteins P-glycoprotein (Pgp) and MRP1 are drug-efflux pumps. In this study, we compared the nucleotide triphosphatase activities of the isolated N-terminal nucleotide binding domains (NBD1) of Pgp and MRP1, and explored the potential role of the phosphorylation target domain of Pgp on the regulation of Pgp NBD1 ATPase activity. We found that: (1) the NBD1s of Pgp and MRP1 have ATPase and GTPase activities, (2) the Kms of Pgp NBD1 for ATP and GTP hydrolysis are identical, while the Km of MRP1 NBD1 for ATP is lower than that for GTP, and (3) phosphorylation of MLD by PKA or PKC produces a marginal increase of Vmax for ATP hydrolysis, without affecting the affinity for ATP. These results show efficient GTP hydrolysis by the NBD1s of Pgp and MRP1, and a minor role of phosphorylation in the control of Pgp NBD1 ATPase activity.

AB - The multidrug resistance proteins P-glycoprotein (Pgp) and MRP1 are drug-efflux pumps. In this study, we compared the nucleotide triphosphatase activities of the isolated N-terminal nucleotide binding domains (NBD1) of Pgp and MRP1, and explored the potential role of the phosphorylation target domain of Pgp on the regulation of Pgp NBD1 ATPase activity. We found that: (1) the NBD1s of Pgp and MRP1 have ATPase and GTPase activities, (2) the Kms of Pgp NBD1 for ATP and GTP hydrolysis are identical, while the Km of MRP1 NBD1 for ATP is lower than that for GTP, and (3) phosphorylation of MLD by PKA or PKC produces a marginal increase of Vmax for ATP hydrolysis, without affecting the affinity for ATP. These results show efficient GTP hydrolysis by the NBD1s of Pgp and MRP1, and a minor role of phosphorylation in the control of Pgp NBD1 ATPase activity.

KW - ABC proteins

KW - CFTR

KW - Expression

KW - Maltose-binding protein

KW - Multidrug resistance

KW - Nucleotide-binding fold

KW - Phosphorylation

KW - Purification

KW - R domain

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U2 - 10.1016/S0006-291X(02)00878-1

DO - 10.1016/S0006-291X(02)00878-1

M3 - Article

C2 - 12163030

AN - SCOPUS:0036389535

SN - 0006-291X

VL - 296

SP - 388

EP - 394

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 2

ER -

Nucleotide triphosphatase activity of the N-terminal nucleotide-binding domains of the multidrug resistance proteins P-glycoprotein and MRP1

Abstract

Keywords

ASJC Scopus subject areas

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