Residues in Na(+) channel D3-S6 segment modulate both batrachotoxin and local anesthetic affinities.

Abstract:

:Batrachotoxin (BTX) alters the gating of voltage-gated Na(+) channels and causes these channels to open persistently, whereas local anesthetics (LAs) block Na(+) conductance. The BTX and LA receptors have been mapped to several common residues in D1-S6 and D4-S6 segments of the Na(+) channel alpha-subunit. We substituted individual residues with lysine in homologous segment D3-S6 of the rat muscle mu1 Na(+) channel from F1274 to N1281 to determine whether additional residues are involved in BTX and LA binding. Two mutant channels, mu1-S1276K and mu1-L1280K, when expressed in mammalian cells, become completely resistant to 5 microM BTX during repetitive pulses. The activation and/or fast inactivation gating of these mutants is substantially different from that of wild type. These mutants also display approximately 10-20-fold reduction in bupivacaine affinity toward their inactivated state but show only approximately twofold affinity changes toward their resting state. These results demonstrate that residues mu1-S1276 and mu1-L1280 in D3-S6 are critical for both BTX and LA binding interactions. We propose that LAs interact readily with these residues from D3-S6 along with those from D1-S6 and D4-S6 in close proximity when the Na(+) channel is in its inactivated state. Implications of this state-dependent binding model for the S6 alignment are discussed.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Wang SY,Nau C,Wang GK

doi

10.1016/S0006-3495(00)76390-9

subject

Has Abstract

pub_date

2000-09-01 00:00:00

pages

1379-87

issue

3

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(00)76390-9

journal_volume

79

pub_type

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