A novel human plasma factor(s) capable of mobilizing intracellular Ca2+ in cultured rat vascular smooth muscle cells.

Using fura-2-loaded rat vascular smooth muscle cells (VSMCs) in culture, we have attempted to partially purify and characterize yet unidentified factor(s) from normal human plasma that stimulates cytoplasmic free Ca2+ concentration ([Ca2+]i). The plasma extract caused an immediate and transient increase of [Ca2+]i in a dose-dependent manner, of which effect was not prevented by pretreatment with either any of receptor antagonists for -adrenergic agonist, angiotensin II, arginine vasopressin, serotonin, thromboxane A2, or with EGTA and nifedipine. This novel plasma factor(s) was heat-stable and completely inactivated by pronase E, suggesting its protein nature. Furthermore, plasma extracts dose-dependently stimulated the accumulation of [3H] inositol phosphates in rat VSMCs. Sephadex G-50 gel chromatography of plasma extracts resolved one major component (mol wt 13,000) and two minor components with larger (greater than 30,000) and smaller (3,000) mol wt. Present study demonstrates the presence of hitherto unidentified plasma factor(s) with size heterogeneity capable of stimulating both mobilization of [Ca2+]i and breakdown of phosphatidylinositol-4,5-biphosphates in rat VSMCs.