SUPRAMOLECULAR ORDER WITHIN THE LENS - H-1-NMR SPECTROSCOPIC EVIDENCE FOR SPECIFIC CRYSTALLIN-CRYSTALLIN INTERACTIONS
SUPRAMOLECULAR ORDER WITHIN THE LENS - H-1-NMR SPECTROSCOPIC EVIDENCE FOR SPECIFIC CRYSTALLIN-CRYSTALLIN INTERACTIONS
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DOI：
10.1006/exer.1994.1146
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发表时间：
1994-11-01
影响因子：
3.4
通讯作者：
CARVER, JA
中科院分区：
文献类型：
--
作者：
COOPER, PG;AQUILINA, JA;CARVER, JA
来自牛晶状体的 α、β 和 γ 晶状体蛋白含有柔性末端延伸，很容易通过 NMR 光谱观察到。因此，通过监测这些共振，核磁共振波谱提供了一种检查晶状体蛋白混合物中特定蛋白质-蛋白质相互作用的方法。本文介绍了牛晶状体核和皮质匀浆以及各种晶状体蛋白混合物的 H-1 NMR 光谱研究。在两种匀浆中，很容易观察到来自 cc-晶状体蛋白的柔性 C 端延伸和 β B2-晶状体蛋白的 N 端延伸的共振，表明这些区域不参与晶状体蛋白-晶状体蛋白相互作用。在皮质匀浆中，还存在来自 γ-晶状体蛋白 N 端短延伸的共振。皮质匀浆比核匀浆产生更强烈的共振，这表明皮质区域具有更多的可移动晶状体蛋白区域。在两种匀浆中，未观察到 β B2-晶状体蛋白的 C 端延伸和 γ(B)-晶状体蛋白的非常短的 C 端延伸。因此，这些蛋白质的 C 末端区域参与与其他晶状体蛋白的相互作用。混合单个晶状体蛋白时观察到类似的效果，例如总 γ-晶状体蛋白和高分子量 β-晶状体蛋白聚集体 (β(H)) 混合物的光谱中不存在 γ(B)-晶状体蛋白的 C 末端延伸。总体而言，结果与晶状体内晶状体蛋白的短程有序一致。
alpha-, beta- and gamma-crystallins from bovine lens contain flexible terminal extensions which are readily observed by NMR spectroscopy. By monitoring these resonances, NMR spectroscopy therefore offers a means of examining specific protein-protein interactions in crystallin mixtures. In this paper, a H-1 NMR spectroscopic study of bovine lens nuclear and cortical homogenates and various crystallin mixtures is presented. In both homogenates, resonances from the flexible C-terminal extensions of cc-crystallin and the N-terminal extension of beta B2-crystallin are readily observed suggesting that these regions are not involved in crystallin-crystallin interactions. In the cortical homogenate, resonances from the short N-terminal extension of gamma(s)-crystallin are also present. The cortical homogenate gives rise to more intense resonances than the nuclear homogenate, suggesting that the cortical region has many more mobile crystallin regions. In both homogenates, the C-terminal extension of beta B2-crystallin and the very short C-terminal extension of gamma(B)-crystallin are not observed. Thus, the C-terminal regions of these proteins are involved in interactions with other crystallins. Similar effects are observed upon mixing of the individual crystallins, e.g. the C-terminal extension of gamma(B)-crystallin is absent in spectra of mixtures of total gamma-crystallin and high-molecular-weight beta-crystallin aggregates (beta(H)). Overall, the results are consistent with a short-range order for the crystallins within the lens.