ABCMdb

ABCC7 p.Lys464Met

ClinVar:	c.1392G>T , p.Lys464Asn ? , not provided
Predicted by SNAP2:	A: D (95%), C: D (95%), D: D (95%), E: D (95%), F: D (95%), G: D (95%), H: D (95%), I: D (95%), L: D (95%), M: D (95%), N: D (91%), P: D (95%), Q: D (95%), R: D (95%), S: D (95%), T: D (95%), V: D (95%), W: D (95%), Y: D (95%),
Predicted by PROVEAN:	A: D, C: D, D: D, E: D, F: D, G: D, H: D, I: D, L: D, M: D, N: D, P: D, Q: D, R: D, S: D, T: D, V: D, W: D, Y: D,

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Publications
[hide] Nucleotide-binding domain 1 of cystic fibrosis tra... Eur J Biochem. 2000 Sep;267(17):5306-12. Duffieux F, Annereau JP, Boucher J, Miclet E, Pamlard O, Schneider M, Stoven V, Lallemand JY
Nucleotide-binding domain 1 of cystic fibrosis transmembrane conductance regulator production of a suitable protein for structural studies.
Eur J Biochem. 2000 Sep;267(17):5306-12., [PMID:10951189]

Abstract [show]
Cystic fibrosis is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR). This protein belongs to the large ATP-binding cassette (ABC) family of transporters. Most patients with cystic fibrosis bear a mutation in the nucleotide-binding domain 1 (NBD1) of CFTR, which plays a key role in the activation of the channel function of CFTR. Determination of the three dimensional structure of NBD1 is essential to better understand its structure-function relationship, and relate it to the biological features of CFTR. In this paper, we report the first preparation of recombinant His-tagged NBD1, as a soluble, stable and isolated domain. The method avoids the use of renaturing processes or fusion constructs. ATPase activity assays show that the recombinant domain is functional. Using tryptophan intrinsic fluorescence, we point out that the local conformation, in the region of the most frequent mutation DeltaF508, could differ from that of the nucleotide-binding subunit of histidine permease, the only available ABC structure. We have undertaken three dimensional structure determination of NBD1, and the first two dimensional 15N-1H NMR spectra demonstrate that the domain is folded. The method should be applicable to the structural studies of NBD2 or of other NBDs from different ABC proteins of major biological interest, such as multidrug resistance protein 1 or multidrug resistance associated protein 1.

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Sentences [show]
No. Sentence Comment
40 Synthetic mutated oligonucleotides (Eurogentec) were K464M: 5H -CCACTGGAGCAGGCATGA- CTTCACTTCTAATGGTG-3H and D572M: 5H -GCTGATTTGT- ATTTATTAATGTCTCCTTTTGGATACC-3H , where underlined nucleotides represent mutations. Login to comment
116 Mutations were introduced in the Walker A and B motifs (K464M, D572M), at positions known to be involved in ATP binding and hydrolysis [21]. Login to comment

[hide] Mutational analysis of the Saccharomyces cerevisia... Mol Microbiol. 1997 Aug;25(4):683-94. Wemmie JA, Moye-Rowley WS
Mutational analysis of the Saccharomyces cerevisiae ATP-binding cassette transporter protein Ycf1p.
Mol Microbiol. 1997 Aug;25(4):683-94., [PMID:9379898]

Abstract [show]
Ycf1p is a member of the ATP-binding cassette transporter family of membrane proteins. Strong sequence similarity has been observed between Ycf1p, the cystic fibrosis transmembrane conductance regulator (CFTR) and multidrug resistance protein (MRP). In this work, we have examined the functional significance of several of the conserved amino acid residues and the genetic requirements for Ycf1p subcellular localization. Biochemical fractionation experiments have established that Ycf1p, expressed at single-copy gene levels, co-fractionates with the vacuolar membrane and that this co-fractionation is independent of vps15, vps34 or end3 gene function. Several cystic fibrosis-associated alleles of the CFTR were introduced into Ycf1p and found to elicit defects analogous to those seen in the CFTR. An amino-terminal extension shared between Ycf1p and MRP, but absent from CFTR, was found to be required for Ycf1p function, but not its subcellular localization. Mutant forms of Ycf1p were also identified that exhibited enhanced biological function relative to the wild-type protein. These studies indicate that Ycf1p will provide a simple, genetically tractable model system for the study of the trafficking and function of ATP-binding cassette transporter proteins, such as the CFTR and MRP.

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133 These mutants corresponded to CFTR alterations known to be associated with cystic fibrosis (G551D and G551S in CFTR, G756D and G756S in Ycf1p) as well as lesions that either disturb normal function (K464M in CFTR, K669M in Ycf1p) or act to suppress the phenotype of ⌬F508 CFTR (R553Q and R553M in CFTR, K758Q and K758M in CFTR). Login to comment
137 The analogous mutations in the CFTR are also non-functional but localized correctly, with the exception of the K464M mutant, which does not function normally and is apparently not properly targeted (Gregory et al., 1991). Login to comment
238 A mutation not associated with CF (K669M in Ycf1p, K464M in CFTR) produced a non-functional, vacuolar membrane-localized protein when present in Ycf1p but was apparently retained in the endoplasmic reticulum in the context of the CFTR (Gregory et al., 1991). Login to comment

[hide] Recombinant synthesis of cystic fibrosis transmemb... Methods Enzymol. 1998;292:686-97. King SA, Sorscher EJ
Recombinant synthesis of cystic fibrosis transmembrane conductance regulator and functional nucleotide-binding domains.
Methods Enzymol. 1998;292:686-97., [PMID:9711592]

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No. Sentence Comment
191 For example, a glycine ~ aspartic acid replacement at CFTR position 551 within a highly conserved ABC protein motif (LSGGQXQR, glycine corresponding to position 551 underlined) disrupts the nucleotide interaction.5 The same appears to be true when a critical lysine residue in the Walker A site of NBD1 is substituted for methionine (K464M.) Login to comment
198 Maximal binding for the wild-type NBD1 was approximately four times that of the G551D and K464M domains. Login to comment

[hide] Defective intracellular transport and processing o... Cell. 1990 Nov 16;63(4):827-34. Cheng SH, Gregory RJ, Marshall J, Paul S, Souza DW, White GA, O'Riordan CR, Smith AE
Defective intracellular transport and processing of CFTR is the molecular basis of most cystic fibrosis.
Cell. 1990 Nov 16;63(4):827-34., [PMID:1699669]

Abstract [show]
The gene associated with cystic fibrosis (CF) encodes a membrane-associated, N-linked glycoprotein called CFTR. Mutations were introduced into CFTR at residues known to be altered in CF chromosomes and in residues believed to play a role in its function. Examination of the various mutant proteins in COS-7 cells indicated that mature, fully glycosylated CFTR was absent from cells containing delta F508, delta 1507, K464M, F508R, and S5491 cDNA plasmids. Instead, an incompletely glycosylated version of the protein was detected. We propose that the mutant versions of CFTR are recognized as abnormal and remain incompletely processed in the endoplasmic reticulum where they are subsequently degraded. Since mutations with this phenotype represent at least 70% of known CF chromosomes, we argue that the molecular basis of most cystic fibrosis is the absence of mature CFTR at the correct cellular location.

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4 Examination of the various mutant proteins in COS-7 cells indicated that mature, fully glycosylated CFTR was absent from cells containing AF508, Al507, K464M, F506R, and S5491 cDNA plasmids. Login to comment
117 Analysis of Mutant Forms of CFTR Expression vectors containing wild-type CFTR (pMT-CFTR, lane 2) and those containing the mutants pMT-CFTR-K464M (lane 3) pMT-CFTR-K1250M (lane 4) pMT-CFTR-Al507 (lane 5) pMT-CFTR-N894. Login to comment
124 K464M cDNA transfected cells, like their Al507 and AF508 nucleotide binding domain 1 mutant counterparts, contain no band C (lane 3). Login to comment
137 CFTR Mutants Mutant CF Exon CFTR Domain A B C Wild type R334W K464M Al507 AF508 F508R s5491 G551 D N894,900Q K1250M Tthllll Y 7 N 9 Y 10 Y 10 N 10 Y 11 Y 11 N 15 N 20 N 22 TM6 NBDl NBDl NBDl NBDl NBDl NBDl ECD4 NBD2 Term - + ++ - + ++ - + - - + - - + - - + - - + - - + ++ + - - - + ++ - + - The known association with CF (Y, yes; N, no), exon localization, domain location, and presence (+ ) or absence (- ) of bands A, B, and C of mutant CFTR species is shown. Login to comment
179 We need to study the ability of these mutants to complement the chloride channel defect in CF epithelial cells to establish, for example, whether K1250M and K464M both abolish function. Login to comment