High-throughput in situ experimental phasing

Acta Crystallogr D Struct Biol. 2020 Aug 1;76(Pt 8):790-801. doi: 10.1107/S2059798320009109. Epub 2020 Jul 28.

Abstract

In this article, a new approach to experimental phasing for macromolecular crystallography (MX) at synchrotrons is introduced and described for the first time. It makes use of automated robotics applied to a multi-crystal framework in which human intervention is reduced to a minimum. Hundreds of samples are automatically soaked in heavy-atom solutions, using a Labcyte Inc. Echo 550 Liquid Handler, in a highly controlled and optimized fashion in order to generate derivatized and isomorphous crystals. Partial data sets obtained on MX beamlines using an in situ setup for data collection are processed with the aim of producing good-quality anomalous signal leading to successful experimental phasing.

Keywords: crystal soaking; experimental phasing; high throughput; in situ crystallography; isomorphism.

MeSH terms

  • Automation, Laboratory* / instrumentation
  • Automation, Laboratory* / methods
  • Crystallography, X-Ray
  • Endopeptidase K / chemistry*
  • Macromolecular Substances / chemistry*
  • Muramidase / chemistry*
  • Synchrotrons / instrumentation

Substances

  • Macromolecular Substances
  • Muramidase
  • Endopeptidase K