The first nucleotide binding fold of the cystic fibrosis transmembrane conductance regulator can function as an active ATPase.

The Journal of Biological Chemistry
Y H Ko, P L Pedersen

Abstract

Cystic fibrosis is caused by mutations in the cell membrane protein called CFTR (cystic fibrosis transmembrane conductance regulator) which functions as a regulated Cl- channel. Although it is known that CFTR contains two nucleotide domains, both of which exhibit the capacity to bind ATP, it has not been demonstrated directly whether one or both domains can function as an active ATPase. To address this question, we have studied the first CFTR nucleotide binding fold (NBF1) in fusion with the maltose-binding protein (MBP), which both stabilizes NBF1 and enhances its solubility. Three different ATPase assays conducted on MBP-NBF1 clearly demonstrate its capacity to catalyze the hydrolysis of ATP. Significantly, the mutations K464H and K464L in the Walker A consensus motif of NBF1 markedly impair its catalytic capacity. MBP alone exhibits no ATPase activity and MBP-NBF1 fails to catalyze the release of phosphate from AMP or ADP. The Vmax of ATP hydrolysis (approximately 30 nmol/min/mg of protein) is significant and is markedly inhibited by azide and by the ATP analogs 2'-(3')-O-(2,4,6-trinitrophenyl)-adenosine-5'-triphosphate and adenosine 5'-(beta, gamma-imido)triphosphate. As inherited mutations within NBF1 account for most case...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Nov 1, 1992·Trends in Genetics : TIG·L C Tsui
Mar 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·K D MacFerrinG L Verdine
Sep 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·K Miwa, M Yoshida
Jan 1, 1987·Methods in Enzymology·T A KunkelR A Zakour
Jan 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·T A Kunkel
May 5, 1995·The Journal of Biological Chemistry·A de Crouy-ChanelG Richarme

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Citations

Oct 6, 2006·Pflügers Archiv : European journal of physiology·Andrei A AleksandrovJohn R Riordan
Feb 5, 2003·Progress in Biophysics and Molecular Biology·Susumu Seino, Takashi Miki
Jul 31, 1998·Trends in Neurosciences·F M Ashcroft, F M Gribble
Jun 1, 1996·Current Opinion in Neurobiology·T J Jentsch
May 22, 2009·Biochemistry·Emily CrowleyRichard Callaghan
Nov 18, 2000·British Journal of Pharmacology·S Trapp, F M Ashcroft
Jul 10, 1999·The Journal of General Physiology·L Csanády, D C Gadsby
Dec 2, 2000·Current Opinion in Pulmonary Medicine·L R Choo-Kang, P L Zeitlin
Mar 11, 2005·Journal of Clinical Gastroenterology·Jonathan A Cohn
Jan 1, 1997·Annual Review of Biochemistry·P D Boyer
Apr 29, 1998·Annual Review of Physiology·J K Foskett
May 9, 2000·Biochemical and Biophysical Research Communications·L D HowellJ A Cohn
Jul 13, 2000·Biochemical and Biophysical Research Communications·A KernA Váradi
Jul 6, 2000·Proceedings of the National Academy of Sciences of the United States of America·M Ikuma, M J Welsh
Jun 17, 1998·Proceedings of the National Academy of Sciences of the United States of America·F M GribbleF M Ashcroft
Aug 5, 1997·Proceedings of the National Academy of Sciences of the United States of America·S TrappF M Ashcroft
Jul 4, 2012·Cellular and Molecular Life Sciences : CMLS·Elena GalfrèOscar Moran
Nov 6, 2004·Gastroenterology Clinics of North America·Jonathan A CohnPaul S Jowell
Mar 8, 2005·Clinics in Laboratory Medicine·Jonathan A CohnPaul S Jowell
Oct 2, 2001·Nihon yakurigaku zasshi. Folia pharmacologica Japonica·A FujitaY Kurachi
Feb 12, 2014·The International Journal of Biochemistry & Cell Biology·Oscar Moran
Dec 2, 2000·The Journal of Biological Chemistry·P E LapinskiM Raghavan
May 24, 2003·Arteriosclerosis, Thrombosis, and Vascular Biology·Roshni R SingarajaMichael R Hayden
Jun 15, 2007·Physiology·Paul M Quinton
Apr 9, 2008·Physiological Reviews·Tsung-Yu Chen, Tzyh-Chang Hwang

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