Document Detail


Coexpression, copurification, crystallization and preliminary X-ray analysis of a complex of ARL2-GTP and PDE delta.
MedLine Citation:
PMID:  11468408     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The small GTPase ARL2 (from Mus musculus) and an effector protein, the delta subunit of human cGMP phosphodiesterase (hPDE delta), were coexpressed and copurified from Escherichia coli as a stable complex. Coexpression significantly increased the otherwise low yield of PDE delta production in E. coli. The complex, which contains ARL2 in the activated GTP-bound form, was crystallized in two forms. The first belongs to the monoclinic space group P2(1), with unit-cell parameters a = 48.1, b = 45.7, c = 74.7 A, beta = 94.0 degrees and one complex (39 kDa) in the asymmetric unit. Cryocooled crystals diffract to 2.3 A using synchrotron radiation. The micro-focused X-ray beam at beamline ID13 (ESRF) allowed the use of very small crystals, which helped to overcome twinning and enabled the identification of a molecular-replacement solution. The second form recrystallized from the first one after several months. These crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 44.5, b = 65.4, c = 104.4 A and one complex in the asymmetric unit. They diffracted to 1.8 A using synchrotron radiation.
Authors:
L Renault; M Hanzal-Bayer; R C Hillig
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2001-07-23
Journal Detail:
Title:  Acta crystallographica. Section D, Biological crystallography     Volume:  57     ISSN:  0907-4449     ISO Abbreviation:  Acta Crystallogr. D Biol. Crystallogr.     Publication Date:  2001 Aug 
Date Detail:
Created Date:  2001-07-24     Completed Date:  2001-10-04     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  9305878     Medline TA:  Acta Crystallogr D Biol Crystallogr     Country:  Denmark    
Other Details:
Languages:  eng     Pagination:  1167-70     Citation Subset:  IM    
Affiliation:
Max-Planck-Institut für Molekulare Physiologie, Abteilung Strukturelle Biologie, Otto-Hahn-Strasse 11, 44227 Dortmund, Germany.
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MeSH Terms
Descriptor/Qualifier:
3',5'-Cyclic-GMP Phosphodiesterases / biosynthesis,  chemistry*,  genetics
Animals
Crystallization
Crystallography, X-Ray
Cyclic Nucleotide Phosphodiesterases, Type 6
Escherichia coli
Eye Proteins / biosynthesis,  chemistry*,  genetics
GTP-Binding Proteins / biosynthesis,  chemistry*,  genetics
Guanosine Triphosphate / chemistry*
Humans
Mice
Protein Conformation
Chemical
Reg. No./Substance:
0/Eye Proteins; 86-01-1/Guanosine Triphosphate; EC 3.1.4.35/3',5'-Cyclic-GMP Phosphodiesterases; EC 3.1.4.35/Cyclic Nucleotide Phosphodiesterases, Type 6; EC 3.1.4.35/PDE6B protein, human; EC 3.1.4.35/Pde6b protein, mouse; EC 3.6.1.-/ARL2 protein, human; EC 3.6.1.-/Arl2 protein, mouse; EC 3.6.1.-/GTP-Binding Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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