Co-crystallization with diabodies: A case study for the introduction of synthetic symmetry

Chelsy Chesterman, Eddy Arnold

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

This work presents a method for introducing synthetic symmetry into protein crystallization samples using an antibody fragment termed a diabody (Dab). These Dabs contain two target binding sites, and engineered disulfide bonds have been included to modulate Dab flexibility. The impacts of Dab engineering have been observed through assessment of thermal stability, small-angle X-ray scattering, and high-resolution crystal structures. Complexes between the engineered Dabs and HIV-1 reverse transcriptase (RT) bound to a high-affinity DNA aptamer were also generated to explore the capacity of engineered Dabs to enable the crystallization of bound target proteins. This strategy increased the crystallization hit frequency obtained for RT-aptamer, and the structure of a Dab-RT-aptamer complex was determined to 3.0-Å resolution. Introduction of synthetic symmetry using a Dab could be a broadly applicable strategy, especially when monoclonal antibodies for a target have previously been identified.

Original languageEnglish (US)
Pages (from-to)598-605.e3
JournalStructure
Volume29
Issue number6
DOIs
StatePublished - Jun 3 2021

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Keywords

  • Dab
  • co-crystallization
  • crystal engineering
  • diabody
  • protein engineering
  • reverse transcriptase
  • synthetic symmetry

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