Busygina O G, Severina I S
Biokhimiia. 1991 Mar;56(3):487-93.
Analysis of soluble guanylate cyclase of rat platelets (105,000 g supernatant) revealed no activating effect of sodium nitroprusside on the enzyme activity. Dithiothreitol (2 x 10(-4) H) added to the sample stimulated the basal activity of guanylate cyclase in the presence of Mg2+ but did not induce the enzyme activation by sodium nitroprusside. Hemoglobin added to the enzyme did not influence its basal activity or the activating effect of sodium nitroprusside. DEAE-Cellulose chromatography of the 105,000 g supernatant revealed two protein peaks, I and II, of which only peak II possessed a guanylate cyclase activity. Fraction I added to a partly purified enzyme did not change the enzyme activity, nor did it enhance the sodium nitroprusside-induced activation of guanylate cyclase. Spectral analysis of the 105,000 g supernatant revealed that the presence of a maximum at 415-425 nm (Soret band) depended on the degree of plasma hemolysis. In the absence of hemolysis the Soret band was unobserved either in the 105,000 g supernatant or in fractions I and II. It is suggested that rat platelet guanylate cyclase is present in these cells in a heme-deficient state.
对大鼠血小板(105,000g 上清液)的可溶性鸟苷酸环化酶进行分析发现,硝普钠对该酶活性无激活作用。向样品中添加二硫苏糖醇(2×10⁻⁴H)在 Mg²⁺存在的情况下刺激了鸟苷酸环化酶的基础活性,但未诱导硝普钠对该酶的激活。向酶中添加血红蛋白不影响其基础活性或硝普钠的激活作用。对 105,000g 上清液进行 DEAE - 纤维素层析显示有两个蛋白峰,I 和 II,其中只有峰 II 具有鸟苷酸环化酶活性。向部分纯化的酶中添加组分 I 既不改变酶活性,也不增强硝普钠诱导的鸟苷酸环化酶激活。对 105,000g 上清液的光谱分析表明,在 415 - 425nm 处(Soret 带)出现最大值取决于血浆溶血程度。在无溶血的情况下,无论是在 105,000g 上清液还是在组分 I 和 II 中均未观察到 Soret 带。提示大鼠血小板鸟苷酸环化酶在这些细胞中以血红素缺乏状态存在。