1XMI

Crystal structure of human F508A NBD1 domain with ATP

Summary for 1XMI

• Entry DOI: 10.2210/pdb1xmi/pdb
• Related: 1Q3H 1R0W 1R0X 1R0Y 1R0Z 1R10 1XMJ
• Descriptor: Cystic fibrosis transmembrane conductance regulator, MAGNESIUM ION, ADENOSINE-5'-TRIPHOSPHATE, ... (4 entities in total)
• Functional Keywords: cftr; nbd1 domain; f508a; cystic fibrosis; nucleotide-binding domain 1, membrane protein, hydrolase
• Biological source: Homo sapiens (human)
• Total number of polymer chains: 5
• Total formula weight: 165046.88

Authors

Lewis, H.A.,Zhao, X.,Wang, C.,Sauder, J.M.,Rooney, I.,Noland, B.W.,Lorimer, D.,Kearins, M.C.,Conners, K.,Condon, B.,Maloney, P.C.,Guggino, W.B.,Hunt, J.F.,Emtage, S.,Structural GenomiX (deposition date: 2004-10-02, release date: 2004-11-09, Last modification date: 2023-08-23)

Primary citation

Lewis, H.A.,Zhao, X.,Wang, C.,Sauder, J.M.,Rooney, I.,Noland, B.W.,Lorimer, D.,Kearins, M.C.,Conners, K.,Condon, B.,Maloney, P.C.,Guggino, W.B.,Hunt, J.F.,Emtage, S.
Impact of the delta-F508 Mutation in First Nucleotide-binding Domain of Human Cystic Fibrosis Transmembrane Conductance Regulator on Domain Folding and Structure
J.Biol.Chem., 280:1346-1353, 2005

PubMed Abstract: Cystic fibrosis is caused by defects in the cystic fibrosis transmembrane conductance regulator (CFTR), commonly the deletion of residue Phe-508 (DeltaF508) in the first nucleotide-binding domain (NBD1), which results in a severe reduction in the population of functional channels at the epithelial cell surface. Previous studies employing incomplete NBD1 domains have attributed this to aberrant folding of DeltaF508 NBD1. We report structural and biophysical studies on complete human NBD1 domains, which fail to demonstrate significant changes of in vitro stability or folding kinetics in the presence or absence of the DeltaF508 mutation. Crystal structures show minimal changes in protein conformation but substantial changes in local surface topography at the site of the mutation, which is located in the region of NBD1 believed to interact with the first membrane spanning domain of CFTR. These results raise the possibility that the primary effect of DeltaF508 is a disruption of proper interdomain interactions at this site in CFTR rather than interference with the folding of NBD1. Interestingly, increases in the stability of NBD1 constructs are observed upon introduction of second-site mutations that suppress the trafficking defect caused by the DeltaF508 mutation, suggesting that these suppressors might function indirectly by improving the folding efficiency of NBD1 in the context of the full-length protein. The human NBD1 structures also solidify the understanding of CFTR regulation by showing that its two protein segments that can be phosphorylated both adopt multiple conformations that modulate access to the ATPase active site and functional interdomain interfaces.

PubMed: 15528182
DOI: 10.1074/jbc.M410968200

Experimental method

X-RAY DIFFRACTION (2.25 Å)