Ito Y, Yoshimoto R, Irimura T, Setaka M, Shimizu H, Osawa T
J Biochem. 1979 Dec;86(6):1807-15. doi: 10.1093/oxfordjournals.jbchem.a132702.
To determine the fluidity of the membrane lipid phase, chicken erythrocytes were labeled with a stearic acid derivative spin label. When chicken erythrocytes were treated with concanavalin A (Con A), ESR spectra showed a change in the peaks of the labels in membrane lipids, indicating an increase of membrane fluidity. The degree of the increase in fluidity of the membrane lipid phase depended on the valency of the lectin used. Tetravalent Con A induced an increase of membrane fluidity at a concentration as low as 30 micrograms/ml, while a monovalent derivative of Con A did not affect membrane fluidity. This increase in membrane fluidity was observed within 10 min after the addition of Con A. If bound Con A was removed with methyl alpha-D-mannoside later than 60 min after its addition, a complete return of the fluidity to the normal level could not be observed. However, no change was found in the composition of phospholipids or in the fatty acid compositions of phosphatidylcholine and phosphatidylethanolamine of chicken erythrocytes after the addition of Con A, indicating that this increase in membrane fluidity is not caused by a change of lipid composition. The clustering of membrane receptors of chicken erythrocytes for Con A was demonstrated when the two-dimensional distribution of ferritin-conjugated Con A on the membranes was assayed by transmission electron microscopy. Furthermore, it was shown that major receptors for Con A of chicken erythrocytes were transmembrane glycoproteins having apparent molecular weights of 100K, 45, and 33K.
为了确定膜脂相的流动性,用硬脂酸衍生物自旋标记物标记鸡红细胞。当用伴刀豆球蛋白A(Con A)处理鸡红细胞时,电子自旋共振(ESR)光谱显示膜脂中标记物的峰发生了变化,表明膜流动性增加。膜脂相流动性增加的程度取决于所用凝集素的价数。四价Con A在低至30微克/毫升的浓度下就能诱导膜流动性增加,而Con A的单价衍生物则不影响膜流动性。在加入Con A后10分钟内就观察到了这种膜流动性的增加。如果在加入Con A 60分钟后再用α-D-甲基甘露糖苷去除结合的Con A,就无法观察到流动性完全恢复到正常水平。然而,加入Con A后,鸡红细胞的磷脂组成或磷脂酰胆碱和磷脂酰乙醇胺的脂肪酸组成没有变化,这表明这种膜流动性的增加不是由脂质组成的变化引起的。当通过透射电子显微镜检测膜上铁蛋白结合的Con A的二维分布时,证明了鸡红细胞Con A膜受体的聚集。此外,还表明鸡红细胞Con A的主要受体是表观分子量为100K、45K和33K的跨膜糖蛋白。