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Input Protein - JBS Floppy-Choppy

JBS Floppy-Choppy is the rescue kit for proteins which are recalcitrant to crystallization. It enables the researcher to modify the protein target by in situ proteolysis to improve its crystallization behavior .

The method implies the addition of trace amounts of protease to the protein solution immediately prior to crystallization. Thus, the crystallization experiment is very straightforward. It can be set up without evaluating the efficacy of proteolysis, without stopping the proteolysis reaction and without purification of any proteolyzed protein fragments.

In situ proteolysis is one of the most efficacious crystallization rescue strategies used at structural genomic centers [1,2].


References

[1] Dong et al. (2007) In situ proteolysis for protein crystallization and structure determination. Nature Methods 4:1019.
[2] Wernimont et al. (2009) In Situ Proteolysis to Generate Crystals for Structure Determination: An Update. PLoS ONE 4:e5094


Selected Literature Citations of JBS Floppy-Choppy

  • Begum et al. (2013) Staphylococcus aureus thiaminase II: oligomerization warrants proteolytic protection against serine proteases Acta Cryst. D 69:2320
  • Faim et al. (2013) Crystallization and preliminary X-ray diffraction analysis of selenophosphate synthetases from Trypanosoma brucei and Leishmania major. Acta Cryst. F 69:864
  • Ochi et al. (2012) Structural Insights into the Role of Domain Flexibility in Human DNA Ligase IV. Structure 20(7):1212
  • Abskharon et al. (2011) Combining in-situ proteolysis and microseed matrix screening to promote crystallization of PrPc-nanobody complexes. PEDS 24(9):737