Cardiac-enriched LIM domain protein fhl2 is required to generate I(Ks) in a heterologous system.

Abstract:

OBJECTIVE:Co-expression of the KvLQT1 and minK potassium channel subunits is required to recapitulate I(Ks), the slow component of the cardiac delayed rectifier current, and mutations in either gene cause the congenital Long QT syndrome. It is becoming increasingly well-recognized that multiprotein channel complexes containing proteins capable of modulating channel function assemble at the plasma membrane. Thus, the aim of our study was to identify proteins involved in I(Ks) modulation. METHODS AND RESULTS:Using a yeast-two-hybrid screen with the intracytoplasmic C-terminus of minK as bait, we identified the cardiac-enriched four-and-a-half LIM domain-containing protein (fhl2) as a potential minK partner. We show interaction between the two proteins in GST pulldown assays and demonstrate overlapping subcellular localization using immunocytochemistry of transfected cells supporting a potential interaction. At the functional level, expression of KvLQT1and minK in HEK cells, which lack endogenous fhl2 protein, generated I(Ks) only when fhl2 was co-expressed. By contrast, in CHO-K1 cells, which express fhl2 endogenously, I(Ks) was suppressed by anti-fhl2 antisense which did not affect the currents generated by KvLQT1alone. CONCLUSION:These data indicate that at least in heterologous cells, the generation of I(Ks) requires fhl2 as an additional protein component.

journal_name

Cardiovasc Res

journal_title

Cardiovascular research

authors

Kupershmidt S,Yang IC,Sutherland M,Wells KS,Yang T,Yang P,Balser JR,Roden DM

doi

10.1016/s0008-6363(02)00498-4

subject

Has Abstract

pub_date

2002-10-01 00:00:00

pages

93-103

issue

1

eissn

0008-6363

issn

1755-3245

pii

S0008636302004984

journal_volume

56

pub_type

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