title: Crystallization of the class IV adenylyl cyclase from Yersinia pestis.
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 A, with one dimer per asymmetric unit and unit-cell parameters a = 33.5, b = 35.5, c = 71.8 A, alpha = 88.7, beta = 82.5, gamma = 65.5 degrees. 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 P2(1)2(1)2(1). These crystals have unit-cell parameters a = 56.8, b = 118.6, c = 144.5 A, diffract to 3 A and probably have two dimers per asymmetric unit and VM = 3.0 A3 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.
authors.affiliation: Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, MD 20899-8310, USA.
authors.forename: Natasha
authors.initials: N
authors.lastname: Smith
authors.forename: Sook-Kyung
authors.initials: SK
authors.lastname: Kim
authors.forename: Prasad T
authors.initials: PT
authors.lastname: Reddy
authors.forename: D Travis
authors.initials: DT
authors.lastname: Gallagher
mesh_terms: D000262:Adenylyl Cyclases / Q000302:isolation & purification*; D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods*; D015010:Yersinia pestis / Q000201:enzymology*
chemical_list: D000262:Adenylyl Cyclases; C406262:adenylyl cyclase 4