Crystallization of the class IV adenylyl cyclase from Yersinia pestis

E-ISSN： 1744-3091|62|3|200-204

ISSN： 1744-3091

Source： Acta Crystallographica Section F, Vol.62, Iss.3, 2006-03, pp. : 200-204

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Abstract

The class IV adenylyl cyclase from Yersinia pestis has been cloned and crystallized in both a triclinic and an orthorhombic form. An amino‐terminal His‐tagged construct, from which the tag was removed by thrombin, crystallized in a triclinic form diffracting to 1.9 Å, with one dimer per asymmetric unit and unit‐cell parameters a = 33.5, b = 35.5, c = 71.8 Å, α = 88.7, β = 82.5, γ = 65.5°. Several mutants of this construct crystallized but diffracted poorly. A non‐His‐tagged native construct (179 amino acids, MW = 20.5 kDa) was purified by conventional chromatography and crystallized in space group P2₁2₁2₁. These crystals have unit‐cell parameters a = 56.8, b = 118.6, c = 144.5 Å, diffract to 3 Å and probably have two dimers per asymmetric unit and V_M = 3.0 Å³ Da⁻¹. Both crystal forms appear to require pH below 5, complicating attempts to incorporate nucleotide ligands into the structure. The native construct has been produced as a selenomethionine derivative and crystallized for phasing and structure determination.