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Orthorhombic lysozyme crystallization at acidic pH values driven by phosphate binding
Author(s) -
Plaza-Garrido Marina,
Salinas-Garcia M. Carmen,
Camara-Artigas Ana
Publication year - 2018
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.374
H-Index - 138
ISSN - 2059-7983
DOI - 10.1107/s205979831800517x
Subject(s) - lysozyme , chemistry , orthorhombic crystal system , crystallography , phosphate , crystallization , sodium , histidine , protein crystallization , inorganic chemistry , crystal structure , amino acid , biochemistry , organic chemistry
The structure of orthorhombic lysozyme has been obtained at 298 K and pH 4.5 using sodium chloride as the precipitant and in the presence of sodium phosphate at a concentration as low as 5 m M . Crystals belonging to space group P 2 1 2 1 2 1 (unit‐cell parameters a = 30, b = 56, c = 73 Å, α = β = γ = 90.00°) diffracted to a resolution higher than 1 Å, and the high quality of these crystals permitted the identification of a phosphate ion bound to Arg14 and His15. The binding of this ion produces long‐range conformational changes affecting the loop containing Ser60–Asn74. The negatively charged phosphate ion shields the electrostatic repulsion of the positively charged arginine and histidine residues, resulting in higher stability of the phosphate‐bound lysozyme. Additionally, a low‐humidity orthorhombic variant was obtained at pH 4.5, and comparison with those previously obtained at pH 6.5 and 9.5 shows a 1.5 Å displacement of the fifth α‐helix towards the active‐site cavity, which might be relevant to protein function. Since lysozyme is broadly used as a model protein in studies related to protein crystallization and amyloid formation, these results indicate that the interaction of some anions must be considered when analysing experiments performed at acidic pH values.