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Deciphering the crystal structure of a novel nanobody against the NEIL1 DNA glycosylase

Acta Crystallogr D Struct Biol. 2024-01; 
Marlo K Thompson, Nidhi Sharma, Andrea Thorn, Aishwarya Prakash
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Molecular Biology Reagents … To optimize expression yields and crystallization, gene blocks containing A5 fused with alternative purification tags (GenScript; Table 1) were inserted into the pEBY vector via standard … Get A Quote

摘要

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 P222 enabled the successful modeling of 96% of residues in the asymmetric unit, with final R and R values of 0.199 and 0.229, respectively... More

关键词

NEIL1 DNA glycosylase, X-ray crystallography, nanobodies, orthorhombic space group, pseudo-merohedral twinning