Crystallization of the class IV adenylyl cyclase from Yersinia pestis.
Acta Crystallogr Sect F Struct Biol Cryst Commun
; 62(Pt 3): 200-4, 2006 Mar 01.
Article
em En
| MEDLINE
| ID: mdl-16511301
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.
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01-internacional
Base de dados:
MEDLINE
Assunto principal:
Yersinia pestis
/
Adenilil Ciclases
/
Cristalização
Idioma:
En
Ano de publicação:
2006
Tipo de documento:
Article