Interaction of bestrophin-1 and Ca2+ channel β-subunits: identification of new binding domains on the bestrophin-1 C-terminus.
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Interaction of bestrophin-1 and Ca2+ channel β-subunits: identification of new binding domains on the bestrophin-1 C-terminus. / Milenkovic, Vladimir M; Krejcova, Sarka; Reichhart, Nadine; Wagner, Andrea; Strauss, Olaf.
In: PLOS ONE, Vol. 6, No. 4, 4, 2011, p. 19364.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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T1 - Interaction of bestrophin-1 and Ca2+ channel β-subunits: identification of new binding domains on the bestrophin-1 C-terminus.
AU - Milenkovic, Vladimir M
AU - Krejcova, Sarka
AU - Reichhart, Nadine
AU - Wagner, Andrea
AU - Strauss, Olaf
PY - 2011
Y1 - 2011
N2 - Bestrophin-1 modulates currents through voltage-dependent L-type Ca(2+) channels by physically interacting with the ?-subunits of Ca(2+) channels. The main function of ?-subunits is to regulate the number of pore-forming Ca(V)-subunits in the cell membrane and modulate Ca(2+) channel currents. To understand the influence of full-length bestrophin-1 on ?-subunit function, we studied binding and localization of bestrophin-1 and Ca(2+) channel subunits, together with modulation of Ca(V)1.3 Ca(2+) channels currents. In heterologeous expression, bestrophin-1 showed co-immunoprecipitation with either, ?3-, or ?4-subunits. We identified a new highly conserved cluster of proline-rich motifs on the bestrophin-1 C-terminus between amino acid position 468 and 486, which enables possible binding to SH3-domains of ?-subunits. A bestrophin-1 that lacks these proline-rich motifs (?CT-PxxP bestrophin-1) showed reduced efficiency to co-immunoprecipitate with ?3 and ?4-subunits. In the presence of ?CT-PxxP bestrophin-1, ?4-subunits and Ca(V)1.3 subunits partly lost membrane localization. Currents from Ca(V)1.3 subunits were modified in the presence of ?4-subunit and wild-type bestrophin-1: accelerated time-dependent activation and reduced current density. With ?CTPxxP bestrophin-1, currents showed the same time-dependent activation as with wild-type bestrophin-1, but the current density was further reduced due to decreased number of Ca(2+) channels proteins in the cell membrane. In summary, we described new proline-rich motifs on bestrophin-1 C-terminus, which help to maintain the ability of ?-subunits to regulate surface expression of pore-forming Ca(V) Ca(2+)-channel subunits.
AB - Bestrophin-1 modulates currents through voltage-dependent L-type Ca(2+) channels by physically interacting with the ?-subunits of Ca(2+) channels. The main function of ?-subunits is to regulate the number of pore-forming Ca(V)-subunits in the cell membrane and modulate Ca(2+) channel currents. To understand the influence of full-length bestrophin-1 on ?-subunit function, we studied binding and localization of bestrophin-1 and Ca(2+) channel subunits, together with modulation of Ca(V)1.3 Ca(2+) channels currents. In heterologeous expression, bestrophin-1 showed co-immunoprecipitation with either, ?3-, or ?4-subunits. We identified a new highly conserved cluster of proline-rich motifs on the bestrophin-1 C-terminus between amino acid position 468 and 486, which enables possible binding to SH3-domains of ?-subunits. A bestrophin-1 that lacks these proline-rich motifs (?CT-PxxP bestrophin-1) showed reduced efficiency to co-immunoprecipitate with ?3 and ?4-subunits. In the presence of ?CT-PxxP bestrophin-1, ?4-subunits and Ca(V)1.3 subunits partly lost membrane localization. Currents from Ca(V)1.3 subunits were modified in the presence of ?4-subunit and wild-type bestrophin-1: accelerated time-dependent activation and reduced current density. With ?CTPxxP bestrophin-1, currents showed the same time-dependent activation as with wild-type bestrophin-1, but the current density was further reduced due to decreased number of Ca(2+) channels proteins in the cell membrane. In summary, we described new proline-rich motifs on bestrophin-1 C-terminus, which help to maintain the ability of ?-subunits to regulate surface expression of pore-forming Ca(V) Ca(2+)-channel subunits.
KW - Animals
KW - Humans
KW - CHO Cells
KW - Cricetinae
KW - Cricetulus
KW - Protein Structure, Secondary
KW - Amino Acid Motifs
KW - Protein Structure, Tertiary
KW - Protein Conformation
KW - Patch-Clamp Techniques
KW - Protein Binding
KW - HEK293 Cells
KW - Microscopy, Confocal/methods
KW - Calcium Channels/chemistry
KW - Chloride Channels/chemistry
KW - Eye Proteins/chemistry
KW - Animals
KW - Humans
KW - CHO Cells
KW - Cricetinae
KW - Cricetulus
KW - Protein Structure, Secondary
KW - Amino Acid Motifs
KW - Protein Structure, Tertiary
KW - Protein Conformation
KW - Patch-Clamp Techniques
KW - Protein Binding
KW - HEK293 Cells
KW - Microscopy, Confocal/methods
KW - Calcium Channels/chemistry
KW - Chloride Channels/chemistry
KW - Eye Proteins/chemistry
U2 - 10.1371/journal.pone.0019364
DO - 10.1371/journal.pone.0019364
M3 - SCORING: Journal article
VL - 6
SP - 19364
JO - PLOS ONE
JF - PLOS ONE
SN - 1932-6203
IS - 4
M1 - 4
ER -
