SUBUNIT 2 (OR BETA) OF RETINAL ROD CGMP-GATED CATION CHANNEL IS A COMPONENT OF THE 240-KDA CHANNEL-ASSOCIATED PROTEIN AND MEDIATES CA2+-CALMODULIN MODULATION

被引:127
作者
CHEN, TY
ILLING, M
MOLDAY, LL
HSU, YT
YAU, KW
MOLDAY, RS
机构
[1] JOHNS HOPKINS UNIV,SCH MED,DEPT NEUROSCI,BALTIMORE,MD 21205
[2] JOHNS HOPKINS UNIV,SCH MED,DEPT OPHTHALMOL,BALTIMORE,MD 21205
[3] JOHNS HOPKINS UNIV,SCH MED,HOWARD HUGHES MED INST,BALTIMORE,MD 21205
[4] UNIV BRITISH COLUMBIA,DEPT BIOCHEM & MOLEC BIOL,VANCOUVER V6T 1Z3,BC,CANADA
关键词
D O I
10.1073/pnas.91.24.11757
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The cGMP-gated cation channel mediating visual transduction in retinal rods was recently found to comprise at least two subunits, 1 and 2 (or alpha and beta). SDS gels of the purified channel show, in addition to a 63-kDa protein band (subunit 1), a 240-kDa protein band that binds Ca2+-calmodulin, a modulator of the channel. To examine any connection between subunit 2 and the 240-kDa protein, cGMP-gated channels formed from the expressed cloned subunits in human embryonic kidney (HEK) 293 cells were tested for Ca2+-calmodulin effect. Homooligomeric channels formed by subunit 1 alone showed no sensitivity to Ca2+-calmodulin, and neither did heterooligomeric channels formed by subunit 1 and the short alternatively spliced form of subunit 2 (2a). By contrast, the cGMP half-activation constant (K-1/2) for hetero-oligomeric channels formed from subunit 1 and the long form of subunit 2 (2b) was increased 1.5- to 2-fold by Ca2+-calmodulin, similar to the increase observed for the native channel. In Western blots of rod outer segment membranes, a subunit 2-specific antibody also recognized the 240-kDa protein. Finally, amino acid sequences derived from peptide fragments of the bovine 240-kDa protein showed approximate to 80% identity to regions of subunit 2b of the human channel. These results together suggest that subunit 2b of the rod channel is a component of the 240-kDa protein and that it mediates the Ca2+-calmodulin modulation of the channel.
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收藏
页码:11757 / 11761
页数:5
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