高分辨率单通道记录在囊性纤维化突变体功能研究中的应用。
Application of high-resolution single-channel recording to functional studies of cystic fibrosis mutants.
作者信息
Cai Zhiwei, Sohma Yoshiro, Bompadre Silvia G, Sheppard David N, Hwang Tzyh-Chang
机构信息
Department of Physiology and Pharmacology, School of Medical Sciences, University of Bristol, Bristol, UK.
出版信息
Methods Mol Biol. 2011;741:419-41. doi: 10.1007/978-1-61779-117-8_27.
Abstract
The patch-clamp technique is a powerful and versatile method to investigate the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel, its malfunction in disease and modulation by small molecules. Here, we discuss how the molecular behaviour of CFTR is investigated using high-resolution single-channel recording and kinetic analyses of channel gating. We review methods used to quantify how cystic fibrosis (CF) mutants perturb the biophysical properties and regulation of CFTR. By explaining the relationship between macroscopic and single-channel currents, we demonstrate how single-channel data provide molecular explanations for changes in CFTR-mediated transepithelial ion transport elicited by CF mutants.
摘要
膜片钳技术是一种强大且通用的方法，用于研究囊性纤维化跨膜传导调节因子（CFTR）氯离子通道、其在疾病中的功能异常以及小分子对其的调节作用。在此，我们讨论如何使用高分辨率单通道记录和通道门控的动力学分析来研究CFTR的分子行为。我们综述了用于量化囊性纤维化（CF）突变体如何扰乱CFTR生物物理特性和调节的方法。通过解释宏观电流和单通道电流之间的关系，我们展示了单通道数据如何为CF突变体引发的CFTR介导的跨上皮离子转运变化提供分子解释。
相似文献
1
Application of high-resolution single-channel recording to functional studies of cystic fibrosis mutants.
Methods Mol Biol. 2011;741:419-41. doi: 10.1007/978-1-61779-117-8_27.
2
Two Small Molecules Restore Stability to a Subpopulation of the Cystic Fibrosis Transmembrane Conductance Regulator with the Predominant Disease-causing Mutation.
J Biol Chem. 2017 Mar 3;292(9):3706-3719. doi: 10.1074/jbc.M116.751537. Epub 2017 Jan 13.
3
Exploiting species differences to understand the CFTR Cl- channel.
Biochem Soc Trans. 2015 Oct;43(5):975-82. doi: 10.1042/BST20150129.
4
Potentiation of the cystic fibrosis transmembrane conductance regulator Cl channel by ivacaftor is temperature independent.
Am J Physiol Lung Cell Mol Physiol. 2018 Nov 1;315(5):L846-L857. doi: 10.1152/ajplung.00235.2018. Epub 2018 Aug 23.
5
Understanding how cystic fibrosis mutations cause a loss of Cl- channel function.
Mol Med Today. 1996 Jul;2(7):290-7. doi: 10.1016/1357-4310(96)10028-9.
6
Differential sensitivity of the cystic fibrosis (CF)-associated mutants G551D and G1349D to potentiators of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel.
J Biol Chem. 2006 Jan 27;281(4):1970-7. doi: 10.1074/jbc.M510576200. Epub 2005 Nov 25.
7
Disease-associated mutations in cytoplasmic loops 1 and 2 of cystic fibrosis transmembrane conductance regulator impede processing or opening of the channel.
Biochemistry. 1997 Sep 30;36(39):11966-74. doi: 10.1021/bi9712652.
8
Impact of the F508del mutation on ovine CFTR, a Cl- channel with enhanced conductance and ATP-dependent gating.
J Physiol. 2015 Jun 1;593(11):2427-46. doi: 10.1113/JP270227. Epub 2015 Apr 9.
9
Inhibition of heterologously expressed cystic fibrosis transmembrane conductance regulator Cl- channels by non-sulphonylurea hypoglycaemic agents.
Br J Pharmacol. 1999 Sep;128(1):108-18. doi: 10.1038/sj.bjp.0702748.
10
Deletion of phenylalanine 508 causes attenuated phosphorylation-dependent activation of CFTR chloride channels.
J Physiol. 2000 May 1;524 Pt 3(Pt 3):637-48. doi: 10.1111/j.1469-7793.2000.00637.x.
引用本文的文献
1
Tuning phenylalanine fluorination to assess aromatic contributions to protein function and stability in cells.
Nat Commun. 2023 Jan 4;14(1):59. doi: 10.1038/s41467-022-35761-w.
2
CFTR bearing variant p.Phe312del exhibits function inconsistent with phenotype and negligible response to ivacaftor.
JCI Insight. 2022 Mar 22;7(6):e148841. doi: 10.1172/jci.insight.148841.
3
Assays of CFTR Function In Vitro, Ex Vivo and In Vivo.
Int J Mol Sci. 2022 Jan 27;23(3):1437. doi: 10.3390/ijms23031437.
4
A small molecule CFTR potentiator restores ATP-dependent channel gating to the cystic fibrosis mutant G551D-CFTR.
Br J Pharmacol. 2022 Apr;179(7):1319-1337. doi: 10.1111/bph.15709. Epub 2022 Jan 21.
5
The gating of the CFTR channel.
Cell Mol Life Sci. 2017 Jan;74(1):85-92. doi: 10.1007/s00018-016-2390-z. Epub 2016 Oct 1.
6
Synergistic Potentiation of Cystic Fibrosis Transmembrane Conductance Regulator Gating by Two Chemically Distinct Potentiators, Ivacaftor (VX-770) and 5-Nitro-2-(3-Phenylpropylamino) Benzoate.
Mol Pharmacol. 2016 Sep;90(3):275-85. doi: 10.1124/mol.116.104570. Epub 2016 Jul 13.
7
Functional reconstitution and channel activity measurements of purified wildtype and mutant CFTR protein.
J Vis Exp. 2015 Mar 9(97):52427. doi: 10.3791/52427.
本文引用的文献
1
Strict coupling between CFTR's catalytic cycle and gating of its Cl- ion pore revealed by distributions of open channel burst durations.
Proc Natl Acad Sci U S A. 2010 Jan 19;107(3):1241-6. doi: 10.1073/pnas.0911061107. Epub 2009 Dec 4.
2
Direct sensing of intracellular pH by the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel.
J Biol Chem. 2009 Dec 18;284(51):35495-506. doi: 10.1074/jbc.M109.072678.
3
State-dependent modulation of CFTR gating by pyrophosphate.
J Gen Physiol. 2009 Apr;133(4):405-19. doi: 10.1085/jgp.200810186.
4
Gating of the CFTR Cl- channel by ATP-driven nucleotide-binding domain dimerisation.
J Physiol. 2009 May 15;587(Pt 10):2151-61. doi: 10.1113/jphysiol.2009.171595. Epub 2009 Mar 30.
5
Potentiation of cystic fibrosis transmembrane conductance regulator (CFTR) Cl- currents by the chemical solvent tetrahydrofuran.
Mol Membr Biol. 2008 Sep;25(6-7):528-38. doi: 10.1080/09687680802487967.
6
Chimeric constructs endow the human CFTR Cl- channel with the gating behavior of murine CFTR.
Proc Natl Acad Sci U S A. 2007 Oct 9;104(41):16365-70. doi: 10.1073/pnas.0701562104. Epub 2007 Oct 3.
7
G551D and G1349D, two CF-associated mutations in the signature sequences of CFTR, exhibit distinct gating defects.
J Gen Physiol. 2007 Apr;129(4):285-98. doi: 10.1085/jgp.200609667. Epub 2007 Mar 12.
8
The two ATP binding sites of cystic fibrosis transmembrane conductance regulator (CFTR) play distinct roles in gating kinetics and energetics.
J Gen Physiol. 2006 Oct;128(4):413-22. doi: 10.1085/jgp.200609622. Epub 2006 Sep 11.
9
F508del CFTR with two altered RXR motifs escapes from ER quality control but its channel activity is thermally sensitive.
Biochim Biophys Acta. 2006 May;1758(5):565-72. doi: 10.1016/j.bbamem.2006.03.006. Epub 2006 Mar 31.
10
The ABC protein turned chloride channel whose failure causes cystic fibrosis.
Nature. 2006 Mar 23;440(7083):477-83. doi: 10.1038/nature04712.
Zotero 插件