Introducing site-specific cysteines into nanobodies for mercury labelling allows de novo phasing of their crystal structures.

ABSTRACT: The generation of high-quality protein crystals and the loss of phase information during an X-ray crystallography diffraction experiment represent the major bottlenecks in the determination of novel protein structures. A generic method for introducing Hg atoms into any crystal independent of the presence of free cysteines in the target protein could considerably facilitate the process of obtaining unbiased experimental phases. Nanobodies (single-domain antibodies) have recently been shown to promote the crystallization and structure determination of flexible proteins and complexes. To extend the usability of nanobodies for crystallographic work, variants of the Nb36 nanobody with a single free cysteine at one of four framework-residue positions were developed. These cysteines could be labe

SUBMITTER: Hansen SB 

PROVIDER: S-EPMC5633906 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature