In situ proteolysis for protein crystallization and structure determination

Nat Methods. 2007 Dec;4(12):1019-21. doi: 10.1038/nmeth1118. Epub 2007 Nov 4.

Abstract

We tested the general applicability of in situ proteolysis to form protein crystals suitable for structure determination by adding a protease (chymotrypsin or trypsin) digestion step to crystallization trials of 55 bacterial and 14 human proteins that had proven recalcitrant to our best efforts at crystallization or structure determination. This is a work in progress; so far we determined structures of 9 bacterial proteins and the human aminoimidazole ribonucleotide synthetase (AIRS) domain.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Crystallization / methods*
  • Crystallography / methods*
  • Peptide Hydrolases / chemistry*
  • Protein Conformation
  • Proteins / chemistry*
  • Proteins / ultrastructure*

Substances

  • Proteins
  • Peptide Hydrolases