Characterization of gadolinium complexes for SAD phasing in macromolecular crystallography: application to CbpF

TY - JOUR

T1 - Characterization of gadolinium complexes for SAD phasing in macromolecular crystallography

T2 - application to CbpF

AU - Molina, Rafael

AU - Stelter, Meike

AU - Kahn, Richard

AU - Hermoso, Juan A

PY - 2009/8

Y1 - 2009/8

N2 - Seven Gd complexes were used in the preparation of heavy-atom derivatives for solving the structure of choline-binding protein F (CbpF), a 36 kDa surface protein from Streptococcus pneumoniae, by the SAD method. CbpF was used as a model system to analyse the phasing capability of each of the derivatives. Three different aspects have been systematically characterized: the efficacy of cocrystallization versus soaking in the binding of the different Gd complexes, their mode of interaction and a comparative study of SAD phasing using synchrotron radiation and using a rotating-anode generator. This study reveals the striking potential of these complexes for SAD phasing using a laboratory source and further reinforces their relevance for high-throughput macromolecular crystallography.

AB - Seven Gd complexes were used in the preparation of heavy-atom derivatives for solving the structure of choline-binding protein F (CbpF), a 36 kDa surface protein from Streptococcus pneumoniae, by the SAD method. CbpF was used as a model system to analyse the phasing capability of each of the derivatives. Three different aspects have been systematically characterized: the efficacy of cocrystallization versus soaking in the binding of the different Gd complexes, their mode of interaction and a comparative study of SAD phasing using synchrotron radiation and using a rotating-anode generator. This study reveals the striking potential of these complexes for SAD phasing using a laboratory source and further reinforces their relevance for high-throughput macromolecular crystallography.

KW - Bacterial Proteins/chemistry

KW - Carrier Proteins/chemistry

KW - Choline/metabolism

KW - Crystallization

KW - Gadolinium/chemistry

KW - Macromolecular Substances/chemistry

KW - Protein Binding

KW - Protein Conformation

KW - Streptococcus pneumoniae

KW - Sulfur/chemistry

KW - Synchrotrons

KW - X-Ray Diffraction

U2 - 10.1107/S0907444909017958

DO - 10.1107/S0907444909017958

M3 - Journal article

C2 - 19622866

SN - 2059-7983

VL - 65

SP - 823

EP - 831

JO - Acta crystallographica. Section D, Biological crystallography

JF - Acta crystallographica. Section D, Biological crystallography

IS - Pt 8

ER -