Processing mutations located throughout the human multidrug resistance P-glycoprotein disrupt interactions between the nucleotide binding domains.

The Journal of Biological Chemistry
Tip W LooDavid M Clarke

Abstract

The most common cause of cystic fibrosis is misfolding of the cystic fibrosis transmembrane conductance regulator (CFTR) protein because of deletion of residue Phe-508 (DeltaF508). P-glycoprotein (P-gp) is an ideal model protein for studying how mutations disrupt folding of ATP-binding cassette proteins such as CFTR because specific chemical chaperones can be used to correct folding defects. Interactions between the nucleotide binding domains (NBDs) are critical because ATP binds at the interface between the NBDs. Here, we used disulfide cross-linking between cysteines in the Walker A sites and the LSGGQ signature sequences to test whether processing mutations located throughout P-gp disrupted interactions between the NBDs. We found that mutations present in the cytoplasmic loops, transmembrane segments, and linker regions or deletion of Tyr-490 (equivalent to Phe-508 in CFTR) inhibited cross-linking between the NBDs. Deletion of Phe-508 in the P-gp/CFTR chimera also inhibited cross-linking between the NBDs. Cross-linking was restored, however, when the mutants were expressed in the presence of the chemical chaperone cyclosporin A. The "rescued" mutants exhibited drug-stimulated ATPase activity, and cross-linking between the NB...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Aug 1, 1992·Nature Genetics·N KartnerJ R Riordan
Jan 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·T A Kunkel
Jan 1, 1982·Methods in Enzymology·C C Goodno
Sep 15, 1995·The Journal of Biological Chemistry·T W Loo, D M Clarke
Aug 18, 1995·The Journal of Biological Chemistry·I L UrbatschA E Senior
Jan 13, 1995·The Journal of Biological Chemistry·T W Loo, D M Clarke
Jan 10, 1997·The Journal of Biological Chemistry·T W Loo, D M Clarke
Apr 18, 1997·The Journal of Biological Chemistry·A A KomarA S Spirin
Jan 31, 1998·The Journal of Biological Chemistry·A N Fedorov, T O Baldwin
Jan 29, 1999·Physiological Reviews·D N Sheppard, M J Welsh
May 20, 1999·Annual Review of Pharmacology and Toxicology·S V AmbudkarM M Gottesman
Oct 3, 1999·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·T W Loo, D M Clarke
Sep 12, 2002·The Journal of Biological Chemistry·Tip W Loo, David M Clarke
Nov 8, 2002·The Journal of Biological Chemistry·Tip W LooDavid M Clarke
Oct 7, 2003·Molecular Cell·Jue ChenFlorante A Quiocho

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Citations

Feb 29, 2008·Annual Review of Biochemistry·John R Riordan
Sep 14, 2011·PloS One·Pedro Eduardo FerreiraJosé Pedro Gil
Jan 31, 2006·FEBS Letters·Annie Frelet, Markus Klein
Jun 16, 2009·Research in Microbiology·Edward N Trifonov
Mar 17, 2009·Biochimica Et Biophysica Acta·King Leung Fung, Michael M Gottesman
Nov 9, 2004·Biochemical and Biophysical Research Communications·Tip W LooDavid M Clarke
May 13, 2006·Journal of Bioenergetics and Biomembranes·Tip W LooDavid M Clarke
May 20, 2015·The Journal of Biological Chemistry·Tip W Loo, David M Clarke

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