Membrane insertion, processing, and topology of cystic fibrosis transmembrane conductance regulator (CFTR) in microsomal membranes

Mingang Chen, Jian Ting Zhang

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-regulated Cl- channel. Malfunction of CFTR causes cystic fibrosis (CF). CFTR belongs to an ATP-binding cassette (ABC) transporter superfamily which includes P-glycoprotein (Pgp), the molecule that is responsible for multidrug resistance in cancer cells. P-glycoprotein molecules have been suggested to have more than one topology and function. In this study, we analysed the early stages of membrane insertion, processing, and topology of human CFTR using rabbit reticulocyte lysate and wheat germ extract translation systems supplemented with canine pancreatic microsomal membranes, Our results suggest that CFTR contains an uncleavable signal sequence and its membrane targeting and insertion may depend on the signal recognition particle (SRP) and SRP receptor. The topology of CFTR in microsomal membranes is the same as the one predicted based on hydropathy plot analysis. These results, together with our previous findings on Pgp, indicate that (1) the topologies of mammalian ABC transporters can be dissected and studied using protein fusion chimeras in a cell-free system; and (2) the membrane targeting and insertion of CFTR and Pgp may take the same pathway, i.e., the SRP dependent pathway, but the membrane folding mechanism of these two proteins in microsomal membranes is probably different.

Original languageEnglish (US)
Pages (from-to)33-40
Number of pages8
JournalMolecular Membrane Biology
Volume13
Issue number1
DOIs
StatePublished - Jan 1 1996

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Membranes
P-Glycoprotein
Signal Recognition Particle
ATP-Binding Cassette Transporters
Cell-Free System
Reticulocytes
Multiple Drug Resistance
Protein Sorting Signals
Cystic Fibrosis
Triticum
Canidae
Proteins
Rabbits

Keywords

  • CFTR
  • In vitro expression
  • P-glycoprotein
  • Signal sequence
  • Topology

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Membrane insertion, processing, and topology of cystic fibrosis transmembrane conductance regulator (CFTR) in microsomal membranes. / Chen, Mingang; Zhang, Jian Ting.

In: Molecular Membrane Biology, Vol. 13, No. 1, 01.01.1996, p. 33-40.

Research output: Contribution to journalArticle

@article{0f9efadb59a74e388dec3a239271adb6,
title = "Membrane insertion, processing, and topology of cystic fibrosis transmembrane conductance regulator (CFTR) in microsomal membranes",
abstract = "Cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-regulated Cl- channel. Malfunction of CFTR causes cystic fibrosis (CF). CFTR belongs to an ATP-binding cassette (ABC) transporter superfamily which includes P-glycoprotein (Pgp), the molecule that is responsible for multidrug resistance in cancer cells. P-glycoprotein molecules have been suggested to have more than one topology and function. In this study, we analysed the early stages of membrane insertion, processing, and topology of human CFTR using rabbit reticulocyte lysate and wheat germ extract translation systems supplemented with canine pancreatic microsomal membranes, Our results suggest that CFTR contains an uncleavable signal sequence and its membrane targeting and insertion may depend on the signal recognition particle (SRP) and SRP receptor. The topology of CFTR in microsomal membranes is the same as the one predicted based on hydropathy plot analysis. These results, together with our previous findings on Pgp, indicate that (1) the topologies of mammalian ABC transporters can be dissected and studied using protein fusion chimeras in a cell-free system; and (2) the membrane targeting and insertion of CFTR and Pgp may take the same pathway, i.e., the SRP dependent pathway, but the membrane folding mechanism of these two proteins in microsomal membranes is probably different.",
keywords = "CFTR, In vitro expression, P-glycoprotein, Signal sequence, Topology",
author = "Mingang Chen and Zhang, {Jian Ting}",
year = "1996",
month = "1",
day = "1",
doi = "10.3109/09687689609160572",
language = "English (US)",
volume = "13",
pages = "33--40",
journal = "Molecular Membrane Biology",
issn = "0968-7688",
publisher = "Informa Healthcare",
number = "1",

}

TY - JOUR

T1 - Membrane insertion, processing, and topology of cystic fibrosis transmembrane conductance regulator (CFTR) in microsomal membranes

AU - Chen, Mingang

AU - Zhang, Jian Ting

PY - 1996/1/1

Y1 - 1996/1/1

N2 - Cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-regulated Cl- channel. Malfunction of CFTR causes cystic fibrosis (CF). CFTR belongs to an ATP-binding cassette (ABC) transporter superfamily which includes P-glycoprotein (Pgp), the molecule that is responsible for multidrug resistance in cancer cells. P-glycoprotein molecules have been suggested to have more than one topology and function. In this study, we analysed the early stages of membrane insertion, processing, and topology of human CFTR using rabbit reticulocyte lysate and wheat germ extract translation systems supplemented with canine pancreatic microsomal membranes, Our results suggest that CFTR contains an uncleavable signal sequence and its membrane targeting and insertion may depend on the signal recognition particle (SRP) and SRP receptor. The topology of CFTR in microsomal membranes is the same as the one predicted based on hydropathy plot analysis. These results, together with our previous findings on Pgp, indicate that (1) the topologies of mammalian ABC transporters can be dissected and studied using protein fusion chimeras in a cell-free system; and (2) the membrane targeting and insertion of CFTR and Pgp may take the same pathway, i.e., the SRP dependent pathway, but the membrane folding mechanism of these two proteins in microsomal membranes is probably different.

AB - Cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-regulated Cl- channel. Malfunction of CFTR causes cystic fibrosis (CF). CFTR belongs to an ATP-binding cassette (ABC) transporter superfamily which includes P-glycoprotein (Pgp), the molecule that is responsible for multidrug resistance in cancer cells. P-glycoprotein molecules have been suggested to have more than one topology and function. In this study, we analysed the early stages of membrane insertion, processing, and topology of human CFTR using rabbit reticulocyte lysate and wheat germ extract translation systems supplemented with canine pancreatic microsomal membranes, Our results suggest that CFTR contains an uncleavable signal sequence and its membrane targeting and insertion may depend on the signal recognition particle (SRP) and SRP receptor. The topology of CFTR in microsomal membranes is the same as the one predicted based on hydropathy plot analysis. These results, together with our previous findings on Pgp, indicate that (1) the topologies of mammalian ABC transporters can be dissected and studied using protein fusion chimeras in a cell-free system; and (2) the membrane targeting and insertion of CFTR and Pgp may take the same pathway, i.e., the SRP dependent pathway, but the membrane folding mechanism of these two proteins in microsomal membranes is probably different.

KW - CFTR

KW - In vitro expression

KW - P-glycoprotein

KW - Signal sequence

KW - Topology

UR - http://www.scopus.com/inward/record.url?scp=0029988492&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029988492&partnerID=8YFLogxK

U2 - 10.3109/09687689609160572

DO - 10.3109/09687689609160572

M3 - Article

C2 - 9147660

AN - SCOPUS:0029988492

VL - 13

SP - 33

EP - 40

JO - Molecular Membrane Biology

JF - Molecular Membrane Biology

SN - 0968-7688

IS - 1

ER -