Diminished self-chaperoning activity of the DeltaF508 mutant of CFTR results in protein misfolding.
The absence of a functional ATP Binding Cassette (ABC) protein called the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) from apical membranes of epithelial cells is responsible for cystic fibrosis (CF). Over 90% of CF patients carry at least one mutant allele with deletion of phenylalan...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2008-02-01
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| Series: | PLoS Computational Biology |
| Online Access: | http://europepmc.org/articles/PMC2265529?pdf=render |
| _version_ | 1819016142201552896 |
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| author | Adrian W R Serohijos Tamás Hegedus John R Riordan Nikolay V Dokholyan |
| author_facet | Adrian W R Serohijos Tamás Hegedus John R Riordan Nikolay V Dokholyan |
| author_sort | Adrian W R Serohijos |
| collection | DOAJ |
| description | The absence of a functional ATP Binding Cassette (ABC) protein called the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) from apical membranes of epithelial cells is responsible for cystic fibrosis (CF). Over 90% of CF patients carry at least one mutant allele with deletion of phenylalanine at position 508 located in the N-terminal nucleotide binding domain (NBD1). Biochemical and cell biological studies show that the DeltaF508 mutant exhibits inefficient biosynthetic maturation and susceptibility to degradation probably due to misfolding of NBD1 and the resultant misassembly of other domains. However, little is known about the direct effect of the Phe508 deletion on the NBD1 folding, which is essential for rational design strategies of cystic fibrosis treatment. Here we show that the deletion of Phe508 alters the folding dynamics and kinetics of NBD1, thus possibly affecting the assembly of the complete CFTR. Using molecular dynamics simulations, we find that meta-stable intermediate states appearing on wild type and mutant folding pathways are populated differently and that their kinetic accessibilities are distinct. The structural basis of the increased misfolding propensity of the DeltaF508 NBD1 mutant is the perturbation of interactions in residue pairs Q493/P574 and F575/F578 found in loop S7-H6. As a proof-of-principle that the S7-H6 loop conformation can modulate the folding kinetics of NBD1, we virtually design rescue mutations in the identified critical interactions to force the S7-H6 loop into the wild type conformation. Two redesigned NBD1-DeltaF508 variants exhibited significantly higher folding probabilities than the original NBD1-DeltaF508, thereby partially rescuing folding ability of the NBD1-DeltaF508 mutant. We propose that these observed defects in folding kinetics of mutant NBD1 may also be modulated by structures separate from the 508 site. The identified structural determinants of increased misfolding propensity of NBD1-DeltaF508 are essential information in correcting this pathogenic mutant. |
| first_indexed | 2024-12-21T02:42:55Z |
| format | Article |
| id | doaj.art-47dbca58aef04f2e950b2187c798b625 |
| institution | Directory Open Access Journal |
| issn | 1553-734X 1553-7358 |
| language | English |
| last_indexed | 2024-12-21T02:42:55Z |
| publishDate | 2008-02-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Computational Biology |
| spelling | doaj.art-47dbca58aef04f2e950b2187c798b6252022-12-21T19:18:38ZengPublic Library of Science (PLoS)PLoS Computational Biology1553-734X1553-73582008-02-0142e100000810.1371/journal.pcbi.1000008Diminished self-chaperoning activity of the DeltaF508 mutant of CFTR results in protein misfolding.Adrian W R SerohijosTamás HegedusJohn R RiordanNikolay V DokholyanThe absence of a functional ATP Binding Cassette (ABC) protein called the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) from apical membranes of epithelial cells is responsible for cystic fibrosis (CF). Over 90% of CF patients carry at least one mutant allele with deletion of phenylalanine at position 508 located in the N-terminal nucleotide binding domain (NBD1). Biochemical and cell biological studies show that the DeltaF508 mutant exhibits inefficient biosynthetic maturation and susceptibility to degradation probably due to misfolding of NBD1 and the resultant misassembly of other domains. However, little is known about the direct effect of the Phe508 deletion on the NBD1 folding, which is essential for rational design strategies of cystic fibrosis treatment. Here we show that the deletion of Phe508 alters the folding dynamics and kinetics of NBD1, thus possibly affecting the assembly of the complete CFTR. Using molecular dynamics simulations, we find that meta-stable intermediate states appearing on wild type and mutant folding pathways are populated differently and that their kinetic accessibilities are distinct. The structural basis of the increased misfolding propensity of the DeltaF508 NBD1 mutant is the perturbation of interactions in residue pairs Q493/P574 and F575/F578 found in loop S7-H6. As a proof-of-principle that the S7-H6 loop conformation can modulate the folding kinetics of NBD1, we virtually design rescue mutations in the identified critical interactions to force the S7-H6 loop into the wild type conformation. Two redesigned NBD1-DeltaF508 variants exhibited significantly higher folding probabilities than the original NBD1-DeltaF508, thereby partially rescuing folding ability of the NBD1-DeltaF508 mutant. We propose that these observed defects in folding kinetics of mutant NBD1 may also be modulated by structures separate from the 508 site. The identified structural determinants of increased misfolding propensity of NBD1-DeltaF508 are essential information in correcting this pathogenic mutant.http://europepmc.org/articles/PMC2265529?pdf=render |
| spellingShingle | Adrian W R Serohijos Tamás Hegedus John R Riordan Nikolay V Dokholyan Diminished self-chaperoning activity of the DeltaF508 mutant of CFTR results in protein misfolding. PLoS Computational Biology |
| title | Diminished self-chaperoning activity of the DeltaF508 mutant of CFTR results in protein misfolding. |
| title_full | Diminished self-chaperoning activity of the DeltaF508 mutant of CFTR results in protein misfolding. |
| title_fullStr | Diminished self-chaperoning activity of the DeltaF508 mutant of CFTR results in protein misfolding. |
| title_full_unstemmed | Diminished self-chaperoning activity of the DeltaF508 mutant of CFTR results in protein misfolding. |
| title_short | Diminished self-chaperoning activity of the DeltaF508 mutant of CFTR results in protein misfolding. |
| title_sort | diminished self chaperoning activity of the deltaf508 mutant of cftr results in protein misfolding |
| url | http://europepmc.org/articles/PMC2265529?pdf=render |
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