Calcium-regulated DNA Binding and Oligomerization of the Neuronal Calcium-sensing Protein, Calsenilin/DREAM/KChIP3

Masanori Osawa, Kit I. Tong, Christina Lilliehook, Wilma Wasco, Joseph D. Buxbaum, H. Y Mary Cheng, Josef M. Penninger, Mitsuhiko Ikura, James B. Ames

Research output: Contribution to journalArticle

107 Citations (Scopus)

Abstract

Calsenilin/DREAM/KChlP3, a member of the recoverin branch of the EF-hand superfamily, interacts with presenilins, serves as a calcium-regulated transcriptional repressor, and interacts with A-type potassium channels. Here we report physicochemical characterization of calcium binding, oligomerization, and DNA binding of human calsenilin/DREAM/KChIP3. Equilibrium Ca2+ binding measurements indicate that the protein binds 3 Ca2+ with a dissociation constant of 14 μM and a Hill coefficient of 0.7. Dynamic light scattering and size exclusion chromatography show that the Ca 2+-bound protein exists as a dimer at protein concentrations lower than 150 μM and forms a tetramer at concentrations above 200 μM. The Ca2+-free protein is a tetramer in the concentration range 20-450 μM. Isothermal titration calorimetry and dynamic light scattering indicate that the Ca2+-free protein tetramer binds endothermically (δH = +25 kcal/mol) to four molecules of DNA derived from the downstream regulatory element (DRE) of either the prodynorphin or c-fos genes. One DRE molecule binds tightly to the protein with a dissociation constant (Kd) of 75 nM, and the other three bind more weakly (Kd = 640 nM). No significant DNA binding was observed for the Ca2+-bound protein. The N-terminal protein fragment (residues 1-70) binds nonspecifically to DRE in a Ca 2+-independent manner, whereas a C-terminal fragment containing the four EF-hands (residues 65-256) binds DRE (Kd = 200 nM) in a Ca 2+-regulated and sequence-specific fashion. The C-terminal fragment is a tetramer in the Ca2+-free state and dissociates into dimers at saturating Ca2+ levels.

Original languageEnglish
Pages (from-to)41005-41013
Number of pages9
JournalJournal of Biological Chemistry
Volume276
Issue number44
DOIs
Publication statusPublished - 2001 Nov 2
Externally publishedYes

Fingerprint

Kv Channel-Interacting Proteins
Oligomerization
Calcium
DNA
Proteins
EF Hand Motifs
Dynamic light scattering
Dimers
Recoverin
Presenilins
fos Genes
Calorimetry
Molecules
Size exclusion chromatography
Potassium Channels
Titration
Gel Chromatography
Genes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Calcium-regulated DNA Binding and Oligomerization of the Neuronal Calcium-sensing Protein, Calsenilin/DREAM/KChIP3. / Osawa, Masanori; Tong, Kit I.; Lilliehook, Christina; Wasco, Wilma; Buxbaum, Joseph D.; Cheng, H. Y Mary; Penninger, Josef M.; Ikura, Mitsuhiko; Ames, James B.

In: Journal of Biological Chemistry, Vol. 276, No. 44, 02.11.2001, p. 41005-41013.

Research output: Contribution to journalArticle

Osawa, M, Tong, KI, Lilliehook, C, Wasco, W, Buxbaum, JD, Cheng, HYM, Penninger, JM, Ikura, M & Ames, JB 2001, 'Calcium-regulated DNA Binding and Oligomerization of the Neuronal Calcium-sensing Protein, Calsenilin/DREAM/KChIP3', Journal of Biological Chemistry, vol. 276, no. 44, pp. 41005-41013. https://doi.org/10.1074/jbc.M105842200
Osawa, Masanori ; Tong, Kit I. ; Lilliehook, Christina ; Wasco, Wilma ; Buxbaum, Joseph D. ; Cheng, H. Y Mary ; Penninger, Josef M. ; Ikura, Mitsuhiko ; Ames, James B. / Calcium-regulated DNA Binding and Oligomerization of the Neuronal Calcium-sensing Protein, Calsenilin/DREAM/KChIP3. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 44. pp. 41005-41013.
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AU - Tong, Kit I.

AU - Lilliehook, Christina

AU - Wasco, Wilma

AU - Buxbaum, Joseph D.

AU - Cheng, H. Y Mary

AU - Penninger, Josef M.

AU - Ikura, Mitsuhiko

AU - Ames, James B.

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