Pieniazek Anna, Gwoździński Krzysztof
Department of Molecular Biophysics University of Łódź, Poland.
Cell Mol Biol Lett. 2003;8(1):127-31.
The effect of the sodium cyanate-induced carbamylation (carbamoylation) of proteins in erythrocytes was studied using spin labelling and spectrophotometric methods. The experiments were conducted in whole blood and in erythrocytes in phosphate buffer using 25 mmol/L of sodium cyanate. Lipid membrane fluidity was determined using three spin-labelled fatty acids: 5-, 12- and 16-doxylstearic acids (5-DS, 12-DS, 16-DS). Internal viscosity was measured with Tempamine, using also EPR spectroscopy. Osmotic fragility was determined spectrophotometrically. Incubation of whole blood with sodium cyanate led to an increase in lipid membrane fluidity in the deeper region of the lipid layer, indicated by 12- and 16-doxylstearic acid, and a decrease near the surface (5-DS). Statistically significant results were obtained for the internal viscosity and osmotic fragility of erythrocytes. An increase in internal viscosity and increase in osmotic fragility were found in erythrocytes after incubation of whole blood, as well as in erythrocytes incubated with sodium cyanate in buffer. Alterations in internal viscosity were stronger in erythrocytes incubated with sodium cyanate in blood than in erythrocytes in the buffer. On the other hand, higher osmotic fragility was observed for erythrocytes in the buffer.
采用自旋标记法和分光光度法研究了氰酸钠诱导的红细胞中蛋白质氨甲酰化作用的影响。实验在全血和磷酸盐缓冲液中的红细胞中进行,使用25 mmol/L的氰酸钠。使用三种自旋标记脂肪酸(5-、12-和16-二氧硬脂酸,即5-DS、12-DS、16-DS)测定脂质膜流动性。使用Tempamine并通过电子顺磁共振光谱法测量内部黏度。通过分光光度法测定渗透脆性。全血与氰酸钠孵育导致脂质层较深区域的脂质膜流动性增加,这由12-和16-二氧硬脂酸表明,而表面附近(5-DS)的流动性降低。红细胞的内部黏度和渗透脆性获得了具有统计学意义的结果。全血孵育后的红细胞以及在缓冲液中与氰酸钠孵育的红细胞中均发现内部黏度增加和渗透脆性增加。与缓冲液中的红细胞相比,血液中与氰酸钠孵育的红细胞内部黏度变化更强。另一方面,缓冲液中的红细胞观察到更高的渗透脆性。