Structure and Dynamics of NBD1 from CFTR Characterized Using Crystallography and Hydrogen/Deuterium Exchange Mass Spectrometry

Lewis, H.A.; Wang, C.; Zhao, Xiaoliang; Hamuro, Yoshitomo; Conners, K.; Kearins, M.C.; Lu, Frances; Sauder, J.M.; Molnar, Kathleen S.; Coales, Stephen J.; Maloney, Peter C.; Guggino, William B.; Wetmore, Diana; Weber, Patricia C; Hunt, J.F.

doi:10.1016/j.jmb.2009.11.051

Cited by 116 publications

(210 citation statements)

References 83 publications

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“…Therefore, a substantial proportion of CF drug-discovery efforts have focused on correction of the molecular defect caused by the F508del mutation. 12,13 F508del is located in the first nucleotide-binding domain (NBD1) of CFTR, [15][16][17][18][19] which is homologous to proteins in the ABC Transporter superfamily. [20][21][22] This name derives from the stereotyped nucleotidebinding domains (NBDs), or ATP-binding cassettes (ABCs), that are conserved among superfamily members.…”

Section: Introductionmentioning

confidence: 99%

“…While CFTR is the only member known to function as an ATP-gated ion channel rather than an ATP-fueled transmembrane pump, its overall domain organization and ATP-dependent mechanochemistry are equivalent to that of ABC Transporters. 4,6,18 These all contain a pair of transmembrane domains (TMDs) that interact with a pair of cytoplasmic ABC domains, which control protein conformation by binding ATP at their mutual interface. [23][24][25] In CFTR, these domains are encoded in a single polypeptide along with a regulatory (R) domain, in the order TMD1, NBD1, R, TMD2, NBD2.…”

Section: Introductionmentioning

confidence: 99%

“…31 However, structural and thermodynamic studies have led to conflicting conclusions regarding the exact molecular defect caused by the F508del mutation. 15,18,19,[32][33][34] Previously published isothermal denaturation studies have failed to reveal a perturbation in the major chemical unfolding transition of human NBD1 (hNBD1). 15,19 Moreover, extensive crystallographic analyses demonstrated that F508del only perturbs the equilibrium structure of Mg-ATP-bound hNBD1 locally at the site of the mutation, which is the region likely to mediate interaction with the transmembrane domains of CFTR.…”

Section: Introductionmentioning

confidence: 99%

“…15,19 Moreover, extensive crystallographic analyses demonstrated that F508del only perturbs the equilibrium structure of Mg-ATP-bound hNBD1 locally at the site of the mutation, which is the region likely to mediate interaction with the transmembrane domains of CFTR. 18 High-resolution hydrogen/deuterium-exchange mass spectrometry analyses of Mg-ATP-bound hNBD1 have measured the influence of the mutation on protein backbone dynamics, which are modestly enhanced near the site of the mutation but not elsewhere in the domain. 18 These results have led to the hypothesis that F508del may interfere with biogenesis by blocking proper interdomain interaction in nascent CFTR 15,35 or by promoting adventitious chaperone interactions near the site of the mutation, 18 rather than by destabilizing hNBD1 itself.…”

Section: Introductionmentioning

confidence: 99%

“…18 High-resolution hydrogen/deuterium-exchange mass spectrometry analyses of Mg-ATP-bound hNBD1 have measured the influence of the mutation on protein backbone dynamics, which are modestly enhanced near the site of the mutation but not elsewhere in the domain. 18 These results have led to the hypothesis that F508del may interfere with biogenesis by blocking proper interdomain interaction in nascent CFTR 15,35 or by promoting adventitious chaperone interactions near the site of the mutation, 18 rather than by destabilizing hNBD1 itself. On the other hand, other results provide evidence that F508del does perturb hNBD1 stability.…”

Section: Introductionmentioning

confidence: 99%

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Integrated biophysical studies implicate partial unfolding of NBD1 of CFTR in the molecular pathogenesis of F508del cystic fibrosis

Wang¹,

Protasevich²,

Yang³

et al. 2010

Protein Science

138

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The lethal genetic disease cystic fibrosis is caused predominantly by in-frame deletion of phenylalanine 508 in the cystic fibrosis transmembrane conductance regulator (CFTR). F508 is located in the first nucleotide-binding domain (NBD1) of CFTR, which functions as an ATP-gated chloride channel on the cell surface. The F508del mutation blocks CFTR export to the surface due to aberrant retention in the endoplasmic reticulum. While it was assumed that F508del interferes with NBD1 folding, biophysical studies of purified NBD1 have given conflicting results concerning the mutation's influence on domain folding and stability. We have conducted isothermal (this paper) and thermal (accompanying paper) denaturation studies of human NBD1 using a variety of biophysical techniques, including simultaneous circular dichroism, intrinsic fluorescence, and static light-scattering measurements. These studies show that, in the absence of ATP, NBD1 unfolds via two sequential conformational transitions. The first, which is strongly influenced by F508del, involves partial unfolding and leads to aggregation accompanied by an increase in tryptophan fluorescence. The second, which is not significantly influenced by F508del, involves full unfolding of NBD1. Mg-ATP binding delays the first transition, thereby offsetting the effect of F508del on domain stability. Evidence suggests that the initial partial unfolding transition is partially responsible for the poor in vitro solubility of human NBD1. Second-site mutations that increase the solubility of isolated F508del-NBD1 in vitro and suppress the trafficking defect of intact F508del-CFTR in vivo also stabilize the protein against this transition, supporting the hypothesize that it is responsible for the pathological trafficking of F508del-CFTR.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Integrated biophysical studies implicate partial unfolding of NBD1 of CFTR in the molecular pathogenesis of F508del cystic fibrosis

Wang¹,

Protasevich²,

Yang³

et al. 2010

Protein Science

138

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Exploring cystic fibrosis using bioinformatics tools: A module designed for the freshman biology course

Zhang¹

2011

Biochem Molecular Bio Educ

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We incorporated a bioinformatics component into the freshman biology course that allows students to explore cystic fibrosis (CF), a common genetic disorder, using bioinformatics tools and skills. Students learn about CF through searching genetic databases, analyzing genetic sequences, and observing the three-dimentional structures of proteins using a number of computational tools. We also use this opportunity to discuss the genetic, molecular, biochemical, and physiological aspects of CF, in an effort to help students appreciate the interrelationships among concepts and topics in biology within a real-world context. This component provides the freshman students at Savannah State University an early exposure to bioinformatics and prepares them for the upper level courses in this area. It also facilitates meaningful and in-depth learning by linking biological concepts and processes through a case study.

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Carbonate‐Promoted Catalytic Activity of Potassium Cations for Soot Combustion by Gaseous Oxygen

et al. 2013

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The possible mechanism of soot combustion catalyzed by potassium carbonate loaded on aluminosilicate was elucidated to understand the surface reaction in solid–solid–gas triphasic catalysis. Potassium species on aluminosilicate showed high catalytic performance for the oxidation of carbon black by gaseous oxygen. Aluminosilicate helped in stabilizing the alkali cation on the surface. The carbonate ion played a critical role in enhancing the catalytic performance by acting as a supplier of active electrons to gaseous oxygen. To the best of our knowledge, this is the first report on the essential role of carbonate on the catalytic activity of an alkali cation for an oxidation reaction, although carbonate (carbon dioxide) is widely recognized to hinder the catalytic performance of the alkali compound for base‐catalyzed reactions.

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Structure and Dynamics of NBD1 from CFTR Characterized Using Crystallography and Hydrogen/Deuterium Exchange Mass Spectrometry

Cited by 116 publications

References 83 publications

Integrated biophysical studies implicate partial unfolding of NBD1 of CFTR in the molecular pathogenesis of F508del cystic fibrosis

Integrated biophysical studies implicate partial unfolding of NBD1 of CFTR in the molecular pathogenesis of F508del cystic fibrosis

Exploring cystic fibrosis using bioinformatics tools: A module designed for the freshman biology course

Carbonate‐Promoted Catalytic Activity of Potassium Cations for Soot Combustion by Gaseous Oxygen

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