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Redox sensitive cysteine residues in calbindin D(28)k are structurally and functionally important

Cedervall, Tommy LU ; Berggård, Tord LU ; Borek, V; Thulin, Eva LU ; Linse, Sara LU and Akerfeldt, KS (2005) In Biochemistry 44(2). p.684-693
Abstract
Human calbindin D-28k is a Ca2+ binding protein that has been implicated in the protection of cells against apoptosis. In this study, the structural and functional significance of the five cysteine residues present in this protein have been investigated through a series of cystein e-to-serine mutations. The mutants were studied under relevant physiological redox potentials in which conformational changes were monitored using ANS binding. Urea-induced denaturations, as monitored by intrinsic tryptophan fluorescence, were also carried out to compare their relative stability. It was shown that the two N-terminal cysteine residues undergo a redox-driven structural change consistent with disulfide bond formation. The other cysteine residues are... (More)
Human calbindin D-28k is a Ca2+ binding protein that has been implicated in the protection of cells against apoptosis. In this study, the structural and functional significance of the five cysteine residues present in this protein have been investigated through a series of cystein e-to-serine mutations. The mutants were studied under relevant physiological redox potentials in which conformational changes were monitored using ANS binding. Urea-induced denaturations, as monitored by intrinsic tryptophan fluorescence, were also carried out to compare their relative stability. It was shown that the two N-terminal cysteine residues undergo a redox-driven structural change consistent with disulfide bond formation. The other cysteine residues are not by themselves sufficient at inducing structural change, but they accentuate the disulfide-dependent conformational change in a redox-dependent manner. Mass spectrometry data show that the three C-terminal cysteine residues can be modified by glutathione. Furthermore, under oxidizing conditions, the data display additional species consistent with the conversion of cysteine thiols to sulfenic acids and disulfides to disulfide-S-monoxides. The biological function of calbindin D-28k appears to be tied to the redox state of the cysteine residues. The two N-terminal cysteine residues are required for activation of myo-inositol monophosphatase, and enzyme activation is enhanced under conditions in which these residues are oxidized. Last, oxidized calbindin D-28k binds Ca2+ with lower affinity than does the reduced protein. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biochemistry
volume
44
issue
2
pages
684 - 693
publisher
The American Chemical Society
external identifiers
  • wos:000226348000027
  • pmid:15641794
  • scopus:12144274839
ISSN
0006-2960
DOI
10.1021/bi049232r
language
English
LU publication?
yes
id
82de11d1-4aaa-4920-ac5c-a31df6a83423 (old id 151574)
date added to LUP
2007-06-29 09:03:30
date last changed
2017-04-16 03:31:44
@article{82de11d1-4aaa-4920-ac5c-a31df6a83423,
  abstract     = {Human calbindin D-28k is a Ca2+ binding protein that has been implicated in the protection of cells against apoptosis. In this study, the structural and functional significance of the five cysteine residues present in this protein have been investigated through a series of cystein e-to-serine mutations. The mutants were studied under relevant physiological redox potentials in which conformational changes were monitored using ANS binding. Urea-induced denaturations, as monitored by intrinsic tryptophan fluorescence, were also carried out to compare their relative stability. It was shown that the two N-terminal cysteine residues undergo a redox-driven structural change consistent with disulfide bond formation. The other cysteine residues are not by themselves sufficient at inducing structural change, but they accentuate the disulfide-dependent conformational change in a redox-dependent manner. Mass spectrometry data show that the three C-terminal cysteine residues can be modified by glutathione. Furthermore, under oxidizing conditions, the data display additional species consistent with the conversion of cysteine thiols to sulfenic acids and disulfides to disulfide-S-monoxides. The biological function of calbindin D-28k appears to be tied to the redox state of the cysteine residues. The two N-terminal cysteine residues are required for activation of myo-inositol monophosphatase, and enzyme activation is enhanced under conditions in which these residues are oxidized. Last, oxidized calbindin D-28k binds Ca2+ with lower affinity than does the reduced protein.},
  author       = {Cedervall, Tommy and Berggård, Tord and Borek, V and Thulin, Eva and Linse, Sara and Akerfeldt, KS},
  issn         = {0006-2960},
  language     = {eng},
  number       = {2},
  pages        = {684--693},
  publisher    = {The American Chemical Society},
  series       = {Biochemistry},
  title        = {Redox sensitive cysteine residues in calbindin D(28)k are structurally and functionally important},
  url          = {http://dx.doi.org/10.1021/bi049232r},
  volume       = {44},
  year         = {2005},
}