Cerami A, Manning J M
Proc Natl Acad Sci U S A. 1971 Jun;68(6):1180-3. doi: 10.1073/pnas.68.6.1180.
The recent use of urea as a treatment for the crisis phase of sickle-cell anemia has prompted us to investigate the possibility that cyanate, which is in equilibrium with urea in solution, might itself prevent the sickling of erythrocytes. We have found that in contrast to the high concentration of urea (1 M) needed to prevent reversibly the in vitro sickling of 80% of the cells, potassium cyanate (0.01-0.10 M) irreversibly inhibits sickling to the same extent. The prevention of sickling is a function of the amount of [(14)C]cyanate incorporated into acidprecipitable protein (0.1-1.0 mol of cyanate per mol of hemoglobin). Most of the radioactivity is accounted for by carbamylation of the NH(2)-terminal valine residues of hemoglobin; there is no detectable carbamylation of the lysine or cysteine residues. The reactive species, HN=C=O (isocyanic acid), may be an analog of O=C=O since both compounds bind to the same valine residues of hemoglobin. Deoxygenated sickled cells also incorporate [(14)C]-cyanate, but the sickling is not reversed. Oxygenation results in normal morphology in 75% of these cells. Upon subsequent deoxygenation, these cells remain normal. Potassium cyanate (5 mM) was also found to be an effective inhibitor of the gelling of deoxyhemoglobin S.
近期使用尿素治疗镰状细胞贫血危象期促使我们研究溶液中与尿素处于平衡状态的氰酸盐自身是否可能阻止红细胞镰变。我们发现,与可逆地阻止80%细胞在体外镰变所需的高浓度尿素(1M)不同,氰酸钾(0.01 - 0.10M)能不可逆地抑制镰变至相同程度。镰变的阻止是掺入酸沉淀蛋白中的[¹⁴C]氰酸盐量的函数(每摩尔血红蛋白含0.1 - 1.0摩尔氰酸盐)。大部分放射性是由血红蛋白氨基末端缬氨酸残基的氨甲酰化引起的;赖氨酸或半胱氨酸残基未检测到氨甲酰化。反应性物质HN = C = O(异氰酸)可能是O = C = O的类似物,因为这两种化合物都与血红蛋白的相同缬氨酸残基结合。脱氧的镰状细胞也掺入[¹⁴C] - 氰酸盐,但镰变未逆转。氧合使这些细胞中的75%恢复正常形态。随后再脱氧时,这些细胞仍保持正常。还发现氰酸钾(5mM)是脱氧血红蛋白S凝胶化的有效抑制剂。