PMID: 2508056Aug 1, 1989Paper

Calmodulin loss in vascular smooth muscle following Triton X-100 or saponin skinning

Pflügers Archiv : European journal of physiology
J P GardnerS R Harris

Abstract

The calmodulin (CaM) content of intact and chemically skinned strips of rat caudal artery was measured using a 125I-CaM radioimmunoassay. The total CaM measured following homogenization of arterial tissue with EGTA and EGTA/Triton X-100 was 2.58 mumol/kg wet tissue. Based on a smooth muscle volume of 40%, this value corresponds to a cellular CaM concentration of 6.5 microM. Approximately 97% of total CaM was soluble and approximately 3% was EGTA-nonextractable. Permeabilization of the plasmalemma with 0.15 mg/ml saponin or 0.5% Triton X-100 caused significant detergent-dependent loss of CaM. At the end of a 1 h skinning period, tissues exposed to saponin lost 30% of total CaM. By comparison, tissues skinned under the same conditions with Triton X-100 lost 50%. During a subsequent 4 h exposure to relaxing solution, total tissue CaM continued to decline. The exponential loss over the 5 h period was described by a first order model having diffusible and nondiffusible CaM components. The diffusible CaM component of saponin skinned tissue (59%) was significantly less than the diffusible component of those skinned with Triton X-100 (88%); however, the rate coefficients for CaM diffusion (0.78 h-1 and 0.91 h-1, respectively) did not s...Continue Reading

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