Expression, purification, crystallization and preliminary X-ray diffraction analysis of a mammalian type 10 adenylyl cyclase.

Kleinboelting, Silke; van den Heuvel, Joop; Kambach, Christian; Weyand, Michael; Leipelt, Martina; Steegborn, Clemens

dc.contributor.author	Kleinboelting, Silke
dc.contributor.author	van den Heuvel, Joop
dc.contributor.author	Kambach, Christian
dc.contributor.author	Weyand, Michael
dc.contributor.author	Leipelt, Martina
dc.contributor.author	Steegborn, Clemens
dc.date.accessioned	2014-09-09T13:41:53Z
dc.date.available	2014-09-09T13:41:53Z
dc.date.issued	2014-04
dc.identifier.citation	Expression, purification, crystallization and preliminary X-ray diffraction analysis of a mammalian type 10 adenylyl cyclase. 2014, 70 (Pt 4):467-9 Acta Crystallogr F Struct Biol Commun	en
dc.identifier.issn	2053-230X
dc.identifier.pmid	24699740
dc.identifier.doi	10.1107/S2053230X14004014
dc.identifier.uri	http://hdl.handle.net/10033/326008
dc.description.abstract	The second messenger cAMP is synthesized in mammals by ten differently regulated adenylyl cyclases (AC1-10). These ACs are grouped into nucleotidyl cyclase class III based on homologies in their catalytic domains. The catalytic domain of AC10 is unique, however, in being activated through direct interaction with calcium and bicarbonate. Here, the production, crystallization and X-ray diffraction analysis of the catalytic domain of human AC10 are described as a basis for structural studies of regulator binding sites and mechanisms. The recombinant protein had high specific AC activity, and crystals of AC10 in space group P63 diffracted to ∼2.0 Å resolution on a synchrotron beamline. A complete diffraction data set revealed unit-cell parameters a = b = 99.65, c = 98.04 Å, indicating one AC10 catalytic domain per asymmetric unit, and confirmed that the obtained crystals are suitable for structure solution and mechanistic studies.
dc.language.iso	en	en
dc.rights	Archived with thanks to Acta crystallographica. Section F, Structural biology communications	en
dc.title	Expression, purification, crystallization and preliminary X-ray diffraction analysis of a mammalian type 10 adenylyl cyclase.	en
dc.type	Article	en
dc.identifier.journal	Acta crystallographica. Section F, Structural biology communications	en
refterms.dateFOA	2018-06-12T22:01:17Z
html.description.abstract	The second messenger cAMP is synthesized in mammals by ten differently regulated adenylyl cyclases (AC1-10). These ACs are grouped into nucleotidyl cyclase class III based on homologies in their catalytic domains. The catalytic domain of AC10 is unique, however, in being activated through direct interaction with calcium and bicarbonate. Here, the production, crystallization and X-ray diffraction analysis of the catalytic domain of human AC10 are described as a basis for structural studies of regulator binding sites and mechanisms. The recombinant protein had high specific AC activity, and crystals of AC10 in space group P63 diffracted to ∼2.0 Å resolution on a synchrotron beamline. A complete diffraction data set revealed unit-cell parameters a = b = 99.65, c = 98.04 Å, indicating one AC10 catalytic domain per asymmetric unit, and confirmed that the obtained crystals are suitable for structure solution and mechanistic studies.