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

5.00
Hdl Handle:
http://hdl.handle.net/10033/326008
Title:
Expression, purification, crystallization and preliminary X-ray diffraction analysis of a mammalian type 10 adenylyl cyclase.
Authors:
Kleinboelting, Silke; van den Heuvel, Joop ( 0000-0001-5085-4010 ) ; Kambach, Christian; Weyand, Michael; Leipelt, Martina; Steegborn, Clemens
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.
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
Journal:
Acta crystallographica. Section F, Structural biology communications
Issue Date:
Apr-2014
URI:
http://hdl.handle.net/10033/326008
DOI:
10.1107/S2053230X14004014
PubMed ID:
24699740
Type:
Article
Language:
en
ISSN:
2053-230X
Appears in Collections:
publications of the research group recombinant protein expression (RPEX)

Full metadata record

DC FieldValue Language
dc.contributor.authorKleinboelting, Silkeen
dc.contributor.authorvan den Heuvel, Joopen
dc.contributor.authorKambach, Christianen
dc.contributor.authorWeyand, Michaelen
dc.contributor.authorLeipelt, Martinaen
dc.contributor.authorSteegborn, Clemensen
dc.date.accessioned2014-09-09T13:41:53Z-
dc.date.available2014-09-09T13:41:53Z-
dc.date.issued2014-04-
dc.identifier.citationExpression, 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 Communen
dc.identifier.issn2053-230X-
dc.identifier.pmid24699740-
dc.identifier.doi10.1107/S2053230X14004014-
dc.identifier.urihttp://hdl.handle.net/10033/326008-
dc.description.abstractThe 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.en
dc.language.isoenen
dc.rightsArchived with thanks to Acta crystallographica. Section F, Structural biology communicationsen
dc.titleExpression, purification, crystallization and preliminary X-ray diffraction analysis of a mammalian type 10 adenylyl cyclase.en
dc.typeArticleen
dc.identifier.journalActa crystallographica. Section F, Structural biology communicationsen

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