The structure of the caspase recruitment domain of BinCARD reveals that all three cysteines can be oxidized

Kai-En Chen, Ayanthi A. Richards, Tom T. Caradoc-Davies, Parimala R. Vajjhala, Gautier Robin, Linda H.L. Lua, Justine M. Hill, Kate Schroder, Matthew J. Sweet, Stuart Kellie, Bostjan Kobe, Jennifer Martin

Research output: Contribution to journalArticleResearchpeer-review

7 Citations (Scopus)

Abstract

The caspase recruitment domain (CARD) is present in death-domain superfamily proteins involved in inflammation and apoptosis. BinCARD is named for its ability to interact with Bcl10 and inhibit downstream signalling. Human BinCARD is expressed as two isoforms that encode the same N-terminal CARD region but which differ considerably in their C-termini. Both isoforms are expressed in immune cells, although BinCARD-2 is much more highly expressed. Crystals of the CARD fold common to both had low symmetry (space group P1). Molecular replacement was unsuccessful in this low-symmetry space group and, as the construct contains no methionines, first one and then two residues were engineered to methionine for MAD phasing. The double-methionine variant was produced as a selenomethionine derivative, which was crystallized and the structure was solved using data measured at two wavelengths. The crystal structures of the native and selenomethionine double mutant were refined to high resolution (1.58 and 1.40 Å resolution, respectively), revealing the presence of a cis-peptide bond between Tyr39 and Pro40. Unexpectedly, the native crystal structure revealed that all three cysteines were oxidized. The mitochondrial localization of BinCARD-2 and the susceptibility of its CARD region to redox modification points to the intriguing possibility of a redox-regulatory role.

Original languageEnglish
Pages (from-to)774-784
Number of pages11
JournalActa Crystallographica Section D: Biological Crystallography
Volume69
Issue number5
DOIs
Publication statusPublished - May 2013
Externally publishedYes

Keywords

  • alternative splicing
  • CARD proteins
  • cysteine modification
  • methionine mutation for MAD phasing
  • mitochondrial localization

Cite this

Chen, Kai-En ; Richards, Ayanthi A. ; Caradoc-Davies, Tom T. ; Vajjhala, Parimala R. ; Robin, Gautier ; Lua, Linda H.L. ; Hill, Justine M. ; Schroder, Kate ; Sweet, Matthew J. ; Kellie, Stuart ; Kobe, Bostjan ; Martin, Jennifer. / The structure of the caspase recruitment domain of BinCARD reveals that all three cysteines can be oxidized. In: Acta Crystallographica Section D: Biological Crystallography. 2013 ; Vol. 69, No. 5. pp. 774-784.
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abstract = "The caspase recruitment domain (CARD) is present in death-domain superfamily proteins involved in inflammation and apoptosis. BinCARD is named for its ability to interact with Bcl10 and inhibit downstream signalling. Human BinCARD is expressed as two isoforms that encode the same N-terminal CARD region but which differ considerably in their C-termini. Both isoforms are expressed in immune cells, although BinCARD-2 is much more highly expressed. Crystals of the CARD fold common to both had low symmetry (space group P1). Molecular replacement was unsuccessful in this low-symmetry space group and, as the construct contains no methionines, first one and then two residues were engineered to methionine for MAD phasing. The double-methionine variant was produced as a selenomethionine derivative, which was crystallized and the structure was solved using data measured at two wavelengths. The crystal structures of the native and selenomethionine double mutant were refined to high resolution (1.58 and 1.40 {\AA} resolution, respectively), revealing the presence of a cis-peptide bond between Tyr39 and Pro40. Unexpectedly, the native crystal structure revealed that all three cysteines were oxidized. The mitochondrial localization of BinCARD-2 and the susceptibility of its CARD region to redox modification points to the intriguing possibility of a redox-regulatory role.",
keywords = "alternative splicing, CARD proteins, cysteine modification, methionine mutation for MAD phasing, mitochondrial localization",
author = "Kai-En Chen and Richards, {Ayanthi A.} and Caradoc-Davies, {Tom T.} and Vajjhala, {Parimala R.} and Gautier Robin and Lua, {Linda H.L.} and Hill, {Justine M.} and Kate Schroder and Sweet, {Matthew J.} and Stuart Kellie and Bostjan Kobe and Jennifer Martin",
year = "2013",
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doi = "10.1107/S0907444913001558",
language = "English",
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pages = "774--784",
journal = "Acta Crystallographica Section D: Structural Biology",
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Chen, K-E, Richards, AA, Caradoc-Davies, TT, Vajjhala, PR, Robin, G, Lua, LHL, Hill, JM, Schroder, K, Sweet, MJ, Kellie, S, Kobe, B & Martin, J 2013, 'The structure of the caspase recruitment domain of BinCARD reveals that all three cysteines can be oxidized', Acta Crystallographica Section D: Biological Crystallography, vol. 69, no. 5, pp. 774-784. https://doi.org/10.1107/S0907444913001558

The structure of the caspase recruitment domain of BinCARD reveals that all three cysteines can be oxidized. / Chen, Kai-En; Richards, Ayanthi A.; Caradoc-Davies, Tom T.; Vajjhala, Parimala R.; Robin, Gautier; Lua, Linda H.L.; Hill, Justine M.; Schroder, Kate; Sweet, Matthew J.; Kellie, Stuart; Kobe, Bostjan; Martin, Jennifer.

In: Acta Crystallographica Section D: Biological Crystallography, Vol. 69, No. 5, 05.2013, p. 774-784.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - The structure of the caspase recruitment domain of BinCARD reveals that all three cysteines can be oxidized

AU - Chen, Kai-En

AU - Richards, Ayanthi A.

AU - Caradoc-Davies, Tom T.

AU - Vajjhala, Parimala R.

AU - Robin, Gautier

AU - Lua, Linda H.L.

AU - Hill, Justine M.

AU - Schroder, Kate

AU - Sweet, Matthew J.

AU - Kellie, Stuart

AU - Kobe, Bostjan

AU - Martin, Jennifer

PY - 2013/5

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N2 - The caspase recruitment domain (CARD) is present in death-domain superfamily proteins involved in inflammation and apoptosis. BinCARD is named for its ability to interact with Bcl10 and inhibit downstream signalling. Human BinCARD is expressed as two isoforms that encode the same N-terminal CARD region but which differ considerably in their C-termini. Both isoforms are expressed in immune cells, although BinCARD-2 is much more highly expressed. Crystals of the CARD fold common to both had low symmetry (space group P1). Molecular replacement was unsuccessful in this low-symmetry space group and, as the construct contains no methionines, first one and then two residues were engineered to methionine for MAD phasing. The double-methionine variant was produced as a selenomethionine derivative, which was crystallized and the structure was solved using data measured at two wavelengths. The crystal structures of the native and selenomethionine double mutant were refined to high resolution (1.58 and 1.40 Å resolution, respectively), revealing the presence of a cis-peptide bond between Tyr39 and Pro40. Unexpectedly, the native crystal structure revealed that all three cysteines were oxidized. The mitochondrial localization of BinCARD-2 and the susceptibility of its CARD region to redox modification points to the intriguing possibility of a redox-regulatory role.

AB - The caspase recruitment domain (CARD) is present in death-domain superfamily proteins involved in inflammation and apoptosis. BinCARD is named for its ability to interact with Bcl10 and inhibit downstream signalling. Human BinCARD is expressed as two isoforms that encode the same N-terminal CARD region but which differ considerably in their C-termini. Both isoforms are expressed in immune cells, although BinCARD-2 is much more highly expressed. Crystals of the CARD fold common to both had low symmetry (space group P1). Molecular replacement was unsuccessful in this low-symmetry space group and, as the construct contains no methionines, first one and then two residues were engineered to methionine for MAD phasing. The double-methionine variant was produced as a selenomethionine derivative, which was crystallized and the structure was solved using data measured at two wavelengths. The crystal structures of the native and selenomethionine double mutant were refined to high resolution (1.58 and 1.40 Å resolution, respectively), revealing the presence of a cis-peptide bond between Tyr39 and Pro40. Unexpectedly, the native crystal structure revealed that all three cysteines were oxidized. The mitochondrial localization of BinCARD-2 and the susceptibility of its CARD region to redox modification points to the intriguing possibility of a redox-regulatory role.

KW - alternative splicing

KW - CARD proteins

KW - cysteine modification

KW - methionine mutation for MAD phasing

KW - mitochondrial localization

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