Gysi J R, Zuber H
Biochem J. 1979 Sep 1;181(3):577-83. doi: 10.1042/bj1810577.
Purified allophycocyanin II and its subunits have been examined with respect to spectroscopic properties, sedimentation, reconstitution and isoelectric behaviour. In 0.02m-potassium phosphate buffer, pH8.0, and at a concentration of 0.25mg/ml, allophycocyanin II and its alpha- and beta-subunits show visible absorption maxima at 650, 615 and 615nm respectively, whereas the fluorescence emission maxima were determined to be at 662, 640 and 630nm respectively. The absorption difference spectrum (dilution difference) of allophycocyanin II displays maxima at 650 and 590nm with a minimum at 610nm. The c.d. spectrum of allophycocyanin II showed only one positive-ellipticity band at 635nm, and a major negative-ellipticity band at 340nm. Oxidation of allophycocyanin II, low- and high-pH solutions (pH3.0 and 11.0), various ethanol concentrations as well as dialysis against distilled water induce a spectral change leading to phycocyanin-like characteristics. In most cases these shifts are reversible. Allophycocyanin II is thermostable over a period of 60min at temperatures up to 60 degrees C. The isoelectric points of allophycocyanin II and its alpha- and beta-subunits are 4.65, 4.64 and 4.82 respectively. Estimated molecular weights from sedimentation-equilibrium analyses were 102500 for allophycocycanin II, 16000 for the alpha- and 31500 for the beta-subunit. Recombination of alpha- and beta-subunits leads to allophycocyanin II, which is indistinguishable from native allophycocyanin with respect to its spectral form, to its gel-filtration and to its electrophoretic behaviour.
已对纯化的别藻蓝蛋白II及其亚基的光谱性质、沉降、重组和等电行为进行了研究。在0.02m磷酸钾缓冲液(pH8.0)中,浓度为0.25mg/ml时,别藻蓝蛋白II及其α-和β-亚基的可见吸收最大值分别在650、615和615nm处,而荧光发射最大值分别测定为662、640和630nm。别藻蓝蛋白II的吸收差光谱(稀释差)在650和590nm处有最大值,在610nm处有最小值。别藻蓝蛋白II的圆二色光谱在635nm处仅显示一个正椭圆率带,在340nm处有一个主要的负椭圆率带。别藻蓝蛋白II在低pH和高pH溶液(pH3.0和11.0)中氧化、不同乙醇浓度以及对蒸馏水透析都会引起光谱变化,导致呈现藻蓝蛋白样特征。在大多数情况下,这些变化是可逆的。别藻蓝蛋白II在高达60℃的温度下60分钟内是热稳定的。别藻蓝蛋白II及其α-和β-亚基的等电点分别为4.65、4.64和4.82。通过沉降平衡分析估计的分子量,别藻蓝蛋白II为102500,α-亚基为16000,β-亚基为31500。α-和β-亚基的重组产生别藻蓝蛋白II,其光谱形式、凝胶过滤和电泳行为与天然别藻蓝蛋白无法区分。