van Haeringen B, Eden D, van den Bogaerde M R, van Grondelle R, Bloemendal M
Dept. of Biophysics, Free University, Amsterdam, The Netherlands.
Eur J Biochem. 1992 Nov 15;210(1):211-6. doi: 10.1111/j.1432-1033.1992.tb17410.x.
Native alpha-crystallin, obtained from the cortex of calf lenses with FPLC (Pharmacia) was characterized by means of transient-electric-birefringence measurements and ultraviolet linear-dichroism spectroscopy. These techniques were also performed on 6-M-urea-dissociated and reconstituted alpha-crystallin. Transient-electric-birefringence measurements offer the possibility to characterize the often observed, but usually neglected, non-spherical occurrences of alpha-crystallin in more detail. Although not distinguishable with size-exclusion chromatography, we could identify at least two different classes of both native and reconstituted alpha-crystallin, from which at least one consists of non-spherical molecules. The results are compared with those obtained with electron microscopy using different staining methods. From the three independent techniques used we find evidence that a fraction of the alpha-crystallin exists in a more extended quaternary structure. The results are difficult to explain with a concentric three-layer model for alpha-crystallin as proposed by Tardieu et al. [Tardieu, A., Laporte, D., Licinio, P., Krop, B. & Delaye, M. (1986) J. Mol. Biol. 192, 711-724].
用快速蛋白质液相色谱法(Pharmacia)从小牛晶状体皮质中获得的天然α-晶状体蛋白,通过瞬态电双折射测量和紫外线性二色光谱进行表征。这些技术也应用于经6M尿素解离和重构的α-晶状体蛋白。瞬态电双折射测量能够更详细地表征α-晶状体蛋白中经常出现但通常被忽视的非球形结构。虽然在尺寸排阻色谱中无法区分,但我们能够识别出至少两种不同类型的天然和重构α-晶状体蛋白,其中至少有一种由非球形分子组成。将结果与使用不同染色方法的电子显微镜观察结果进行比较。从所使用的三种独立技术中,我们发现有证据表明一部分α-晶状体蛋白以更伸展的四级结构存在。这些结果难以用Tardieu等人 [Tardieu, A., Laporte, D., Licinio, P., Krop, B. & Delaye, M. (1986) J. Mol. Biol. 192, 711 - 724] 提出的α-晶状体蛋白同心三层模型来解释。