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

Rafael Molina; Meike Stelter; Richard Kahn; Juan A Hermoso

doi:10.1107/S0907444909017958

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

Rafael Molina, Meike Stelter, Richard Kahn, Juan A Hermoso

Protein Structure and Function Program

9 Citationer (Scopus)

Abstract

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.

Originalsprog	Engelsk
Tidsskrift	Acta crystallographica. Section D, Biological crystallography
Vol/bind	65
Udgave nummer	Pt 8
Sider (fra-til)	823-31
Antal sider	9
ISSN	2059-7983
DOI	https://doi.org/10.1107/S0907444909017958
Status	Udgivet - aug. 2009

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10.1107/S0907444909017958

Citationsformater

@article{5e3f4e8f08074a449c16bfde7b0c8574,

title = "Characterization of gadolinium complexes for SAD phasing in macromolecular crystallography: application to CbpF",

abstract = "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.",

keywords = "Bacterial Proteins/chemistry, Carrier Proteins/chemistry, Choline/metabolism, Crystallization, Gadolinium/chemistry, Macromolecular Substances/chemistry, Protein Binding, Protein Conformation, Streptococcus pneumoniae, Sulfur/chemistry, Synchrotrons, X-Ray Diffraction",

author = "Rafael Molina and Meike Stelter and Richard Kahn and Hermoso, {Juan A}",

year = "2009",

month = aug,

doi = "10.1107/S0907444909017958",

language = "English",

volume = "65",

pages = "823--31",

journal = "Acta Crystallographica Section D: Structural Biology",

issn = "2059-7983",

publisher = "International Union of Crystallography",

number = "Pt 8",

}

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: Structural Biology

JF - Acta Crystallographica Section D: Structural Biology

IS - Pt 8

ER -

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

Abstract

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