Marlo K Thompson, Nidhi Sharma, Andrea Thorn, Aishwarya Prakash
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引用次数: 0
摘要
纳米抗体（VHHs）是具有三个抗原CDR区的单域抗体，可用于多种科学应用。在这里，我们结晶了一种 14 kDa 的纳米抗体（A5），它特异于参与碱基切除修复途径第一步的内切酶 VIII (Nei)-like 1 或 NEIL1 DNA 糖基化酶，并测定了其 2.1 Å 分辨率的结构。晶体可能存在孪晶和各向异性衍射，这给研究带来了挑战。尽管孪晶指标不确定，但在正交环境中进行再处理并在空间群 P21212 中进行分子置换，成功地对不对称单元中 96% 的残基进行了建模，最终的 Rwork 值和 Rfree 值分别为 0.199 和 0.229。
Deciphering the crystal structure of a novel nanobody against the NEIL1 DNA glycosylase.
Nanobodies (VHHs) are single-domain antibodies with three antigenic CDR regions and are used in diverse scientific applications. Here, an ∼14 kDa nanobody (A5) specific for the endonuclease VIII (Nei)-like 1 or NEIL1 DNA glycosylase involved in the first step of the base-excision repair pathway was crystallized and its structure was determined to 2.1 Å resolution. The crystals posed challenges due to potential twinning and anisotropic diffraction. Despite inconclusive twinning indicators, reprocessing in an orthorhombic setting and molecular replacement in space group P21212 enabled the successful modeling of 96% of residues in the asymmetric unit, with final Rwork and Rfree values of 0.199 and 0.229, respectively.
期刊介绍：
Acta Crystallographica Section D welcomes the submission of articles covering any aspect of structural biology, with a particular emphasis on the structures of biological macromolecules or the methods used to determine them.
Reports on new structures of biological importance may address the smallest macromolecules to the largest complex molecular machines. These structures may have been determined using any structural biology technique including crystallography, NMR, cryoEM and/or other techniques. The key criterion is that such articles must present significant new insights into biological, chemical or medical sciences. The inclusion of complementary data that support the conclusions drawn from the structural studies (such as binding studies, mass spectrometry, enzyme assays, or analysis of mutants or other modified forms of biological macromolecule) is encouraged.
Methods articles may include new approaches to any aspect of biological structure determination or structure analysis but will only be accepted where they focus on new methods that are demonstrated to be of general applicability and importance to structural biology. Articles describing particularly difficult problems in structural biology are also welcomed, if the analysis would provide useful insights to others facing similar problems.
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