Open AccessCrystallization of the class IV adenylyl cyclase from Yersinia pestis
Author(s) -
Smith Natasha,
Kim SookKyung,
Reddy Prasad T.,
Gallagher D. Travis
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106002855
Subject(s) - yersinia pestis , adenylyl cyclase , microbiology and biotechnology , crystallization , virology , chemistry , medicine , biology , enzyme , biochemistry , virulence , gene , organic chemistry
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 P 2 1 2 1 2 1 . 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 Å 3 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.