Institute of Experimental Cardiology, Russian Cardiology Research and Production Complex, Moscow, 121552, Russia.
Biochemistry (Mosc). 2015 Mar;80(3):284-95. doi: 10.1134/S0006297915030049.
A 3D-model of bovine testicular hyaluronidase (BTH) was constructed based on established tertiary structure of human hyaluronidase Hyal1 using a molecular homological modeling method in silico. The analysis of the BTH 3D-model demonstrated lysine residue stratification during enzyme modification. The 3D-model of chondroitin sulfate (CHS)-modified hyaluronidase (BTH-CHS) was obtained by modeling covalent binding of lysine residues with benzoquinone-activated CHS. The degree of enzyme modification and the length of CHS chains were varied during 3D modeling. The importance of deep BTH modification degree for the formation of active and stable enzyme derivatives was shown, as determined earlier experimentally. The effective size of the CHS coat for productive BTH modification was confirmed. It is theoretically achieved at the increase in molecular mass of BTH-CHS derivative to approximately 140-180 kDa and can be practically obtained, according to experimental data, using CHS of different molecular mass (30-50 as well as 120-140 kDa).
基于人透明质酸酶 Hyal1 的已建立的三级结构，使用计算机分子同源建模方法构建了牛睾丸透明质酸酶（BTH）的 3D 模型。BTH 3D 模型分析表明，在酶修饰过程中存在赖氨酸残基分层。通过建模赖氨酸残基与苯醌激活的硫酸软骨素（CHS）的共价结合，获得了硫酸软骨素（CHS）修饰的透明质酸酶（BTH-CHS）的 3D 模型。在 3D 建模过程中，改变了酶修饰的程度和 CHS 链的长度。如早期实验所示，显示了深 BTH 修饰程度对于形成活性和稳定的酶衍生物的重要性。证实了 CHS 涂层的有效大小对于生产性 BTH 修饰是重要的。根据实验数据，理论上可以在 BTH-CHS 衍生物的分子量增加到约 140-180 kDa 时达到，并且可以使用不同分子量的 CHS（30-50 以及 120-140 kDa）实际获得。
