Getting the best out of long-wavelength X-rays: de novo chlorine/sulfur SAD phasing of a structural protein from ATV

TY - JOUR

T1 - Getting the best out of long-wavelength X-rays: de novo chlorine/sulfur SAD phasing of a structural protein from ATV

AU - Goulet, Adeline

AU - Vestergaard, Gisle Alberg

AU - Felisberto-Rodrigues, Catarina

AU - Campanacci, Valérie

AU - Garrett, Roger Antony

AU - Cambillau, Christian

AU - Ortiz-Lombardía, Miguel

PY - 2010/2/12

Y1 - 2010/2/12

N2 - The structure of a 14 kDa structural protein from Acidianus two-tailed virus (ATV) was solved by single-wavelength anomalous diffraction (SAD) phasing using X-ray data collected at 2.0 Å wavelength. Although the anomalous signal from methionine sulfurs was expected to suffice to solve the structure, one chloride ion turned out to be essential to achieve phasing. The minimal data requirements and the relative contributions of the Cl and S atoms to phasing are discussed. This work supports the feasibility of a systematic approach for the solution of protein crystal structures by SAD based on intrinsic protein light atoms along with associated chloride ions from the solvent. In such cases, data collection at long wavelengths may be a time-efficient alternative to seleno-methionine substitution and heavy-atom derivatization.

AB - The structure of a 14 kDa structural protein from Acidianus two-tailed virus (ATV) was solved by single-wavelength anomalous diffraction (SAD) phasing using X-ray data collected at 2.0 Å wavelength. Although the anomalous signal from methionine sulfurs was expected to suffice to solve the structure, one chloride ion turned out to be essential to achieve phasing. The minimal data requirements and the relative contributions of the Cl and S atoms to phasing are discussed. This work supports the feasibility of a systematic approach for the solution of protein crystal structures by SAD based on intrinsic protein light atoms along with associated chloride ions from the solvent. In such cases, data collection at long wavelengths may be a time-efficient alternative to seleno-methionine substitution and heavy-atom derivatization.

KW - Acidianus

KW - Chlorine

KW - Crystallography, X-Ray

KW - Models, Molecular

KW - Protein Structure, Tertiary

KW - Selenomethionine

KW - Sulfur

KW - Viral Structural Proteins

KW - X-Rays

U2 - 10.1107/s0907444909051798

DO - 10.1107/s0907444909051798

M3 - Journal article

C2 - 20179342

SN - 2059-7983

VL - 66

SP - 304

EP - 308

JO - Acta Crystallographica Section D: Structural Biology

JF - Acta Crystallographica Section D: Structural Biology

IS - Pt 3

ER -