The inositol 1,4,5-trisphosphate-gated Ca2+ channel: Effect of the protein thiol reagent thimerosal on channel activity

The solubilized partially purified Ins(1,4,5)P-3-sensitive Ca2+ channel from rat cerebellum has been reconstituted into planar lipid bilayer membranes by the 'tip-dip' method [Ehrlich (1992) Methods Enzymol. 207, 463-471] allowing low noise current records. Single-channel events have been recorded. In the presence of 10 mu M Ins(1,4,5)P-3, 50 mu M ATP, and 0.2 mu M Ca2+ the Ins(1,4,5)P-3 receptor channel opens to a conductance level of 53 pS. In the presence of 100 mu M thimerosal (TMS), a sulphydryl-oxidizing agent, three subconductance levels (60 pS, 80 pS and 120 pS) were observed. More than one population of mean open times was found, both in the absence and presence of TMS, although TMS affected the length of the open time by decreasing the short opening significantly from 4.05 ms to 2.78 ms and increasing the longer open time from 27.8 ms to 94.8 ms. The results indicate that TMS enhances Ins(1,4,5)P-3-induced Ca2+ release by both altering the open times of the channel significantly and causing a shift to higher subconductance levels.