Blockade of complement activation in bullous pemphigoid by using recombinant CD55-CD46 fusion protein.
Blockade of complement activation in bullous pemphigoid by using recombinant CD55-CD46 fusion protein.
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DOI：
10.1097/cm9.0000000000001312
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发表时间：
2021-04-05
影响因子：
6.1
通讯作者：
Dang EL
中科院分区：
文献类型：
--
作者：
Qiao P;Luo YX;Zhi DL;Wang G;Dang EL
致编辑：大疱性类天疱疮（BP）是一种自身免疫性水疱性疾病，由自身抗体引起，并与补体激活有关。[1]自身抗体介导的补体激活参与了BP的发病机制。补体调节蛋白（CRP）包括CD 35、CD 46、CD 55和CD 59，它们是补体系统中一类重要的调节蛋白，控制酶级联反应、膜攻击复合物的组装和补体系统稳态。[2]通常，CD 55和CD 46通过抑制新的C3和C5转化酶的产生并加速已经形成的那些转化酶的降解来负调节补体系统活化，从而保护细胞免受补体活化诱导的损伤。迄今为止，不同CRP的分子融合已被用于产生具有新功能的嵌合分子。[3]本研究利用ClustalW（version 2.1，UCD，爱尔兰）、Vector NTI Viewer（version 4.0，UCD. 1，BioScience Technology reader，USA）和国家生物技术信息中心（GenBank编号：NM_000574. 5，NM_002389。4）。两种蛋白质具有相似的整体结构，包括一个信号肽，四个补体控制蛋白重复结构域和一个富含丝氨酸/苏氨酸的结构域。为了实现对BP的更大抑制作用，将两种蛋白质的编码序列框内连接以产生编码重组DM融合蛋白的嵌合基因[图1A]。将融合基因通过Nde I和Xho I酶切位点连接到pET-28 a表达载体中，得到图1B所示的重组质粒;引物位点的引物P1序列为50-ccaccatatggactgtggccttcccccagat-30（Nde I酶切位点加下划线），
To the Editor: Bullous pemphigoid (BP) is an autoimmune blistering disease that is caused by autoantibodies and is associated with complement activation.[1] Autoantibodymediated complement activation is involved in BP pathogenesis. Complement-regulatory proteins (CRPs) including CD35, CD46, CD55, and CD59 comprise an important class of regulatory proteins in the complement system that control the enzyme cascades, assembly of the membraneattack complex, and complement system homeostasis.[2] Classically, both CD55 and CD46 negatively regulate complement system activation by inhibiting the production of new C3 and C5 convertases and accelerating the degradation of those already formed, thereby protecting cells from complement activation-induced damage. To date, the molecular fusion of different CRPs has been used to create chimeric molecules with novel functions.[3] In this study, we generated a novel decay-accelerating factor and membrane cofactor protein (DAF-MCP, CD55-CD46) fusion protein to achieve a higher inhibitory activity against BP.The amino acid and nucleotide sequences of CD55 and CD46 were analyzed and codon-optimized using ClustalW (version 2.1, UCD, Ireland), Vector NTI Viewer (version 4.0. 1, BioScience Technology readers, USA) and National Center for Biotechnology Information (GenBank number: NM_000574. 5, NM_002389. 4). Both proteins have similar overall structures comprising one signal peptide, four complement control protein-repeat domains, and one serine/threonine-rich domain. To achieve greater inhibitory effects for BP, the coding sequences of both proteins were joined in-frame to produce a chimeric gene encoding the recombinant DM fusion protein [Figure 1A]. The fusion gene was ligated into the pET-28a expression vector through the Nde I and Xho I restriction sites to obtain the recombinant plasmid shown in Figure 1B; the primer P1 sequence for the primer site was 50-ccaccatatggactgtggccttcccccagat-30 (Nde I restriction site underlined),