Referenced in: none

Automatically associated channels: Kv10.1

Title: Effects of sulfhydryl reagents on [3H] inositol trisphosphate binding to dog cerebellar membranes.

Authors: C M Yang, H C Lee

Journal, date & volume: J. Recept. Res., 1989 , 9, 159-69

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/2545875

Abstract
Homogenates from dog cerebellum were fractionated using sucrose gradient centrifugation. The [3H]inositol 1,4,5-trisphosphate binding and the glucose 6-phosphatase activities were found to co-purify. The binding was saturable and had high affinity (Bmax = 44 pmol/mg protein, Kd = 116 nM). Selective chemical modification was used to examine amino acid residues of the microsomal receptor that might be critical for the binding of inositol trisphophate. Sulfhydryl reagents, p-chloromercuricphenyl sulfonic acid. eosin 5-maleimide, N-ethyl maleimide and fluorescein 5-maleimide were found to be highly potent inhibitors of the binding with half-maximal inhibition occurring at about 20 microM, 70 microM, 1 mM, and 0.1 mM, respectively. The inhibition was specific since the presence of 10 microM of inositol trisphosphate during the reaction completely protected against the inhibition by these reagents. These results suggest that sulfhydryl group is essential for inositol trisphosphate binding to its receptor.