Odintsova E S, Dmitrenok P S, Baranova S V, Timofeeva A M, Buneva V N, Nevinsky G A
Institute of Chemical Biology and Fundamental Medicine, Siberian Division of the Russian Academy of Sciences, Novosibirsk, 630090, Russia.
Biochemistry (Mosc). 2015 Feb;80(2):180-201. doi: 10.1134/S0006297915020054.
It was shown previously that, as differentiated from canonical proteases, abzymes against myelin basic protein (MBP) from blood of patients with multiple sclerosis and systemic lupus erythematosus effectively cleaved only MBP, while antibodies (ABs) against integrase (IN) from blood of HIV-infected patients specifically hydrolyzed only IN. In this work, all sites of effective hydrolysis by anti-IN antibodies (IgG and IgM) of 25-mer oligopeptide (OP25) corresponding to MBP were identified using reversed-phase and thin-layer chromatographies and MALDI mass spectrometry. It was found that amino acid sequences of OP25 and other oligopeptides hydrolyzed by anti-MBP abzymes were partially homologous to some fragments of the full sequence of IN. Sequences of IN oligopeptides cleavable by anti-IN abzymes were homologous to some fragments of MBP, but anti-MBP abzymes could not effectively hydrolyze OPs corresponding to IN. The common features of the cleavage sites of OP25 and other oligopeptides hydrolyzed by anti-MBP and anti-IN abzymes were revealed. The literature data on hydrolysis of specific and nonspecific proteins and oligopeptides by abzymes against different protein antigens were analyzed. Overall, the literature data suggest that short OPs, including OP25, mainly interact with light chains of polyclonal ABs, which had lower affinity and specificity to the substrate than intact ABs. However, it seems that anti-IN ABs are the only one example of abzymes capable of hydrolyzing various oligopeptides with high efficiency (within some hours but not days). Possible reasons for the efficient hydrolysis of foreign oligopeptides by anti-IN abzymes from HIV-infected patients are discussed.
先前的研究表明，与典型蛋白酶不同，来自多发性硬化症和系统性红斑狼疮患者血液中的抗髓鞘碱性蛋白（MBP）抗体酶仅能有效切割MBP，而来自HIV感染患者血液中的抗整合酶（IN）抗体则仅能特异性水解IN。在这项工作中，使用反相色谱、薄层色谱和基质辅助激光解吸电离质谱法（MALDI）确定了与MBP对应的25聚体寡肽（OP25）被抗IN抗体（IgG和IgM）有效水解的所有位点。研究发现，OP25和其他被抗MBP抗体酶水解的寡肽的氨基酸序列与IN全序列的某些片段部分同源。可被抗IN抗体酶切割的IN寡肽序列与MBP的某些片段同源，但抗MBP抗体酶不能有效水解与IN对应的寡肽。揭示了抗MBP和抗IN抗体酶水解OP25和其他寡肽的切割位点的共同特征。分析了关于针对不同蛋白质抗原的抗体酶水解特异性和非特异性蛋白质及寡肽的文献数据。总体而言，文献数据表明，包括OP25在内的短寡肽主要与多克隆抗体的轻链相互作用，其对底物的亲和力和特异性低于完整抗体。然而，似乎抗IN抗体是能够高效水解各种寡肽的抗体酶的唯一例子（在数小时内而非数天内）。讨论了HIV感染患者的抗IN抗体酶高效水解外来寡肽的可能原因。
