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Ca 2+ activation of hSlo K + channel is suppressed by N‐terminal GFP tag
Author(s) -
Meyer Elisabeth,
Fromherz Peter
Publication year - 1999
Publication title -
european journal of neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.346
H-Index - 206
eISSN - 1460-9568
pISSN - 0953-816X
DOI - 10.1046/j.1460-9568.1999.00548.x
Subject(s) - terminal (telecommunication) , chemistry , channel (broadcasting) , green fluorescent protein , microbiology and biotechnology , biology , gene , computer network , computer science , biochemistry
The human slow poke (hSlo) K + channel was tagged with GFP (green fluorescent protein) at the N‐terminus of its α‐subunit. The fusion protein was expressed transiently in HEK293 cells; it formed functional voltage‐gated channels as shown by whole cell patch‐clamp measurements. However, the tag lowered the voltage dependence of gating and it suppressed the typical left‐shift of gating by intracellular binding of Ca 2+ . The location of the GFP‐tagged N‐terminus was confirmed to be on the extracellular side by application of a monoclonal antibody to nonpermeabilized cells. Structural interpretations of the effects are discussed.