当前位置：
X-MOL 学术
›
Biochemistry
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Translating the Concept of Bispecific Antibodies to Engineering Heterodimeric Phosphotriesterases with Broad Organophosphate Substrate Recognition
Biochemistry ( IF 2.9 ) Pub Date : 2020-11-04 , DOI: 10.1021/acs.biochem.0c00751 Benjamin Escher 1 , Anja Köhler 1, 2 , Laura Job 1 , Franz Worek 2 , Arne Skerra 1
Biochemistry ( IF 2.9 ) Pub Date : 2020-11-04 , DOI: 10.1021/acs.biochem.0c00751 Benjamin Escher 1 , Anja Köhler 1, 2 , Laura Job 1 , Franz Worek 2 , Arne Skerra 1
Affiliation
|
We have adopted the concept of bispecific antibodies, which can simultaneously block or cross-link two different biomolecular targets, to create bispecific enzymes by exploiting the homodimeric quaternary structure of bacterial phosphotriesterases (PTEs). The PTEs from Brevundimonas diminuta and Agrobacterium radiobacter, whose engineered variants can efficiently hydrolyze organophosphorus (OP) nerve agents and pesticides, respectively, have attracted considerable interest for the treatment of the corresponding intoxications. OP nerve agents and pesticides still pose a severe toxicological threat in military conflicts, including acts of terrorism, as well as in agriculture, leading to >100000 deaths per year. In principle, engineered conventional homodimeric PTEs may provoke hydrolytic inactivation of individual OPs in vivo, and their application as catalytic bioscavengers via administration into the bloodstream has been proposed. However, their narrow substrate specificity would necessitate therapeutic application of a set or mixture of different enzymes, which complicates biopharmaceutical development. We succeeded in combining subunits from both enzymes and to stabilize their heterodimerization by rationally designing electrostatic steering mutations, thus breaking the natural C2 symmetry. The resulting bispecific enzyme from two PTEs with different bacterial origin exhibits an ultrabroad OP substrate profile and allows the efficient detoxification of both nerve agents and pesticides. Our approach of combining two active sites with distinct substrate specificities within one artificial dimeric biocatalyst—retaining the size and general properties of the original enzyme without utilizing protein mixtures or much larger fusion proteins—not only should facilitate biological drug development but also may be applicable to oligomeric enzymes with other catalytic activities.
中文翻译：
将双特异性抗体的概念转化为具有广泛的有机磷酸酯底物识别功能的工程异二聚磷酸三酯酶。
我们采用了双特异性抗体的概念，该双特异性抗体可以同时阻断或交联两个不同的生物分子靶标，从而通过利用细菌磷酸三酯酶（PTE）的同二聚体四级结构来创建双特异性酶。短杆菌Bremundimonas diminuta和放射土壤杆菌的PTE，其工程化的变体可以分别有效水解有机磷（OP）神经毒剂和杀虫剂，在治疗相应的中毒方面引起了极大的兴趣。OP神经毒剂和农药在包括恐怖主义行为在内的军事冲突以及农业中仍然构成严重的毒理学威胁，每年导致超过100000人死亡。原则上，工程改造的常规同二聚体PTE可能在体内引起单个OP的水解失活并提出了通过将其施用到血液中作为催化生物清除剂的应用。然而，它们狭窄的底物特异性将需要治疗性应用一组或多种不同酶的混合物，这使生物药物开发复杂化。我们成功地结合了两种酶的亚基，并通过合理设计静电操纵突变来稳定它们的异二聚化作用，从而打破了自然的C2对称性。由具有不同细菌来源的两个PTE产生的双特异性酶表现出超宽的OP底物谱，并能对神经毒剂和农药有效地排毒。
更新日期：2020-11-17
中文翻译：
将双特异性抗体的概念转化为具有广泛的有机磷酸酯底物识别功能的工程异二聚磷酸三酯酶。
我们采用了双特异性抗体的概念，该双特异性抗体可以同时阻断或交联两个不同的生物分子靶标，从而通过利用细菌磷酸三酯酶（PTE）的同二聚体四级结构来创建双特异性酶。短杆菌Bremundimonas diminuta和放射土壤杆菌的PTE，其工程化的变体可以分别有效水解有机磷（OP）神经毒剂和杀虫剂，在治疗相应的中毒方面引起了极大的兴趣。OP神经毒剂和农药在包括恐怖主义行为在内的军事冲突以及农业中仍然构成严重的毒理学威胁，每年导致超过100000人死亡。原则上，工程改造的常规同二聚体PTE可能在体内引起单个OP的水解失活并提出了通过将其施用到血液中作为催化生物清除剂的应用。然而，它们狭窄的底物特异性将需要治疗性应用一组或多种不同酶的混合物，这使生物药物开发复杂化。我们成功地结合了两种酶的亚基，并通过合理设计静电操纵突变来稳定它们的异二聚化作用，从而打破了自然的C2对称性。由具有不同细菌来源的两个PTE产生的双特异性酶表现出超宽的OP底物谱，并能对神经毒剂和农药有效地排毒。
京公网安备 11010802027423号