Domain organization of murine mdr1b P-glycoprotein: The cytoplasmic linker region is a potential dimerization domain

Shailaja Rao Juvvadi, Joseph S. Glavy, Susan Band Horwitz, George A. Orr

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

P-glycoprotein is an integral membrane protein that functions as a cytotoxic drug-efflux pump. Studies suggest that the transporter exists in the membrane as a dimer and possibly higher order structures. We report the bacterial expression of the linker region (amino acids 621-688) of murine mdr1b P-glycoprotein and demonstrate that this region, which serves to link the two homologous halves of the transporter, has the potential to serve as a dimerization domain. The recombinant peptide (8742 daltons) eluted from a gel filtration column at a position corresponding to a dimer (i.e. 17,500 daltons). A dimer:monomer equilibrium, as a function of peptide concentration, was confirmed by large zone gel filtration chromatography. The dissociation constant (K(D)) for the dimer:monomer equilibrium was 350 nM. It was possible to dissociate the dimer by low pH (3.0) or high ionic strength (2.5 M NaCl). Dimerization was not affected by an alkaline pH of 10 or 5 mM EDTA. Studies with a truncated linker peptide indicated that the N-terminal 48 amino acids were sufficient for dimerization.

Original languageEnglish (US)
Pages (from-to)442-447
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume230
Issue number2
DOIs
StatePublished - Jan 13 1997

Fingerprint

Dimerization
P-Glycoprotein
Dimers
Peptides
Gel Chromatography
Amino Acids
Monomers
Gels
Edetic Acid
Osmolar Concentration
Membrane Proteins
Chromatography
Ionic strength
Membranes
Pumps
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Domain organization of murine mdr1b P-glycoprotein : The cytoplasmic linker region is a potential dimerization domain. / Juvvadi, Shailaja Rao; Glavy, Joseph S.; Band Horwitz, Susan; Orr, George A.

In: Biochemical and Biophysical Research Communications, Vol. 230, No. 2, 13.01.1997, p. 442-447.

Research output: Contribution to journalArticle

@article{140608307ae441d4abef787ad7e93cb4,
title = "Domain organization of murine mdr1b P-glycoprotein: The cytoplasmic linker region is a potential dimerization domain",
abstract = "P-glycoprotein is an integral membrane protein that functions as a cytotoxic drug-efflux pump. Studies suggest that the transporter exists in the membrane as a dimer and possibly higher order structures. We report the bacterial expression of the linker region (amino acids 621-688) of murine mdr1b P-glycoprotein and demonstrate that this region, which serves to link the two homologous halves of the transporter, has the potential to serve as a dimerization domain. The recombinant peptide (8742 daltons) eluted from a gel filtration column at a position corresponding to a dimer (i.e. 17,500 daltons). A dimer:monomer equilibrium, as a function of peptide concentration, was confirmed by large zone gel filtration chromatography. The dissociation constant (K(D)) for the dimer:monomer equilibrium was 350 nM. It was possible to dissociate the dimer by low pH (3.0) or high ionic strength (2.5 M NaCl). Dimerization was not affected by an alkaline pH of 10 or 5 mM EDTA. Studies with a truncated linker peptide indicated that the N-terminal 48 amino acids were sufficient for dimerization.",
author = "Juvvadi, {Shailaja Rao} and Glavy, {Joseph S.} and {Band Horwitz}, Susan and Orr, {George A.}",
year = "1997",
month = "1",
day = "13",
doi = "10.1006/bbrc.1996.5932",
language = "English (US)",
volume = "230",
pages = "442--447",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Domain organization of murine mdr1b P-glycoprotein

T2 - The cytoplasmic linker region is a potential dimerization domain

AU - Juvvadi, Shailaja Rao

AU - Glavy, Joseph S.

AU - Band Horwitz, Susan

AU - Orr, George A.

PY - 1997/1/13

Y1 - 1997/1/13

N2 - P-glycoprotein is an integral membrane protein that functions as a cytotoxic drug-efflux pump. Studies suggest that the transporter exists in the membrane as a dimer and possibly higher order structures. We report the bacterial expression of the linker region (amino acids 621-688) of murine mdr1b P-glycoprotein and demonstrate that this region, which serves to link the two homologous halves of the transporter, has the potential to serve as a dimerization domain. The recombinant peptide (8742 daltons) eluted from a gel filtration column at a position corresponding to a dimer (i.e. 17,500 daltons). A dimer:monomer equilibrium, as a function of peptide concentration, was confirmed by large zone gel filtration chromatography. The dissociation constant (K(D)) for the dimer:monomer equilibrium was 350 nM. It was possible to dissociate the dimer by low pH (3.0) or high ionic strength (2.5 M NaCl). Dimerization was not affected by an alkaline pH of 10 or 5 mM EDTA. Studies with a truncated linker peptide indicated that the N-terminal 48 amino acids were sufficient for dimerization.

AB - P-glycoprotein is an integral membrane protein that functions as a cytotoxic drug-efflux pump. Studies suggest that the transporter exists in the membrane as a dimer and possibly higher order structures. We report the bacterial expression of the linker region (amino acids 621-688) of murine mdr1b P-glycoprotein and demonstrate that this region, which serves to link the two homologous halves of the transporter, has the potential to serve as a dimerization domain. The recombinant peptide (8742 daltons) eluted from a gel filtration column at a position corresponding to a dimer (i.e. 17,500 daltons). A dimer:monomer equilibrium, as a function of peptide concentration, was confirmed by large zone gel filtration chromatography. The dissociation constant (K(D)) for the dimer:monomer equilibrium was 350 nM. It was possible to dissociate the dimer by low pH (3.0) or high ionic strength (2.5 M NaCl). Dimerization was not affected by an alkaline pH of 10 or 5 mM EDTA. Studies with a truncated linker peptide indicated that the N-terminal 48 amino acids were sufficient for dimerization.

UR - http://www.scopus.com/inward/record.url?scp=0031566187&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031566187&partnerID=8YFLogxK

U2 - 10.1006/bbrc.1996.5932

DO - 10.1006/bbrc.1996.5932

M3 - Article

C2 - 9016799

AN - SCOPUS:0031566187

VL - 230

SP - 442

EP - 447

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 2

ER -