Radiation-induced site-specific damage of mercury derivatives

Phasing and implications

Udupi A. Ramagopal, Zbigniew Dauter, Radhakannan Thirumuruhan, Elena Fedorov, Steven C. Almo

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

The behavior of mercury-derivatized triclinic crystals of a 60 kDa protein target from the New York Structural GenomiX Research Consortium provides novel insights into the mechanism of heavy-atom-specific radiation damage and its potential exploitation for de novo structure solution. Despite significant anomalous signal, structure solution by classic SAD and MAD phasing approaches was not successful. A detailed analysis revealed that significant isomorphic variation of the diffracted intensities was induced by X-ray irradiation. These intensity changes allowed the crystal structure to be solved by the radiation-damage-induced phasing (RIP) technique. Inspection of the crystal structure and electron-density maps demonstrated that the covalent S-Hg bonds at all four derivatized cysteine sites were much more susceptible to radiation-induced cleavage than other bonds typically present in native proteins. A simple diagnostic is described to identify the fingerprint of such decay at the time of data collection/processing. The rapid radiation-induced decomposition of mercury adducts is consistent with the difficulties frequently associated with the experimental phasing of mercury derivatives and suggests a straightforward solution to overcome this limitation by radiation-damage-induced phasing with anomalous scattering (RIPAS). These results indicate that historically recalcitrant and newly emerging difficulties associated with Hg phasing should be revisited.

Original languageEnglish (US)
Pages (from-to)1289-1298
Number of pages10
JournalActa Crystallographica Section D: Biological Crystallography
Volume61
Issue number9
DOIs
StatePublished - Sep 2005

Fingerprint

Radiation damage
Mercury
radiation damage
Radiation
damage
Derivatives
radiation
Crystal structure
proteins
crystal structure
cysteine
exploitation
adducts
Carrier concentration
Cysteine
inspection
cleavage
emerging
Proteins
Inspection

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics
  • Condensed Matter Physics
  • Structural Biology

Cite this

Radiation-induced site-specific damage of mercury derivatives : Phasing and implications. / Ramagopal, Udupi A.; Dauter, Zbigniew; Thirumuruhan, Radhakannan; Fedorov, Elena; Almo, Steven C.

In: Acta Crystallographica Section D: Biological Crystallography, Vol. 61, No. 9, 09.2005, p. 1289-1298.

Research output: Contribution to journalArticle

Ramagopal, Udupi A. ; Dauter, Zbigniew ; Thirumuruhan, Radhakannan ; Fedorov, Elena ; Almo, Steven C. / Radiation-induced site-specific damage of mercury derivatives : Phasing and implications. In: Acta Crystallographica Section D: Biological Crystallography. 2005 ; Vol. 61, No. 9. pp. 1289-1298.
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