Purification, crystallization and preliminary X-ray analysis of a maize cytokinin glucoside specific beta-glucosidase

Abstrakt

Zm-p60.1, a cytokinin glucoside specific beta -glucosidase from maize, is a key enzyme involved in plant development and growth. It has been overexpressed in soluble form from Escherichia coli with a His tag at its N-terminus. The recombinant protein has been purified and crystallized at room temperature using PEG 4000 as the main precipitant. At least three crystal forms have been observed from very similar growth conditions. A flash-annealed monoclinic crystal diffracted to high resolution (beyond 2 Angstrom) with space group P2(1) and unit-cell parameters a = 55.66, b = 110.72, c = 72.94 Angstrom, beta = 92.10 degrees. The asymmetric unit is estimated and confirmed by molecular-replacement solution to contain one Zm-p60.1 dimer, giving a crystal volume per protein mass (V-M) of 1.89 Angstrom (3) Da(-1) and a solvent content of 35%.