PMID: 9529383May 16, 1998Paper

Dissection of de novo membrane insertion activities of internal transmembrane segments of ATP-binding-cassette transporters: toward understanding topological rules for membrane assembly of polytopic membrane proteins

Molecular Biology of the Cell
J T ZhangC Wang

Abstract

The membrane assembly of polytopic membrane proteins is a complicated process. Using Chinese hamster P-glycoprotein (Pgp) as a model protein, we investigated this process previously and found that Pgp expresses more than one topology. One of the variations occurs at the transmembrane (TM) domain including TM3 and TM4: TM4 inserts into membranes in an N(in)-C(out) rather than the predicted N(out)-C(in) orientation, and TM3 is in cytoplasm rather than the predicted N(in)-C(out) orientation in the membrane. It is possible that TM4 has a strong activity to initiate the N(in)-C(out) membrane insertion, leaving TM3 out of the membrane. Here, we tested this hypothesis by expressing TM3 and TM4 in isolated conditions. Our results show that TM3 of Pgp does not have de novo N(in)-C(out) membrane insertion activity whereas TM4 initiates the N(in)-C(out) membrane insertion regardless of the presence of TM3. In contrast, TM3 and TM4 of another polytopic membrane protein, cystic fibrosis transmembrane conductance regulator (CFTR), have a similar level of de novo Nin-Cout membrane insertion activity and TM4 of CFTR functions only as a stop-transfer sequence in the presence of TM3. Based on these findings, we propose that 1) the membrane inser...Continue Reading

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Citations

Jun 25, 2003·The Journal of Membrane Biology·R J Turner
Jun 30, 2000·Current Opinion in Cell Biology·R E DalbeyJ C Samuelson
Mar 8, 2000·Microbiology and Molecular Biology Reviews : MMBR·M van Geest, J S Lolkema
Feb 27, 1999·Current Opinion in Structural Biology·P J Booth, A R Curran
Feb 9, 2012·Journal of Molecular Biology·Stella A NeugebauerSandra J Facey
Jun 14, 2002·FEBS Letters·Imogen CoeZlatina Naydenova
Oct 17, 1998·The Journal of Biological Chemistry·K OtaK Mihara
Feb 25, 2009·Journal of Molecular Biology·Karl EnquistIngMarie Nilsson

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