Crystallization of the class IV adenylyl cyclase from Yersinia pestis.

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TitleCrystallization of the class IV adenylyl cyclase from Yersinia pestis.
Publication TypeJournal Article
Year of Publication2006
AuthorsSmith, N, Kim, S-K, Reddy, PT, D Gallagher, T
JournalActa Crystallogr Sect F Struct Biol Cryst Commun
Volume62
IssuePt 3
Pagination200-4
Date Published2006 Mar 1
ISSN1744-3091
KeywordsAdenylate Cyclase, Cloning, Molecular, Crystallization, 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.

DOI10.1107/S1744309106002855
Alternate JournalActa Crystallogr. Sect. F Struct. Biol. Cryst. Commun.
PubMed ID16511301
PubMed Central IDPMC2197185