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Specificity Analysis of Protein Lysine Methyltransferases Using SPOT Peptide Arrays
Published on: November 29, 2014
Lysine methylation as a routine rescue strategy for protein crystallization
Thomas S Walter1, Christoph Meier, Rene Assenberg
1Oxford Protein Production Facility, The Henry Wellcome Building for Genomic Medicine, Oxford University, Roosevelt Drive, Oxford OX3 7BN, United Kingdom.
Structure (London, England : 1993)
|November 14, 2006
Summary
Chemical modification by reductive methylation of lysine residues can improve protein crystallization for X-ray structure determination. This simple method successfully crystallized refractory proteins, yielding diffraction-quality crystals and structures.
Area of Science:
- Structural Biology
- Protein Crystallization
- Biochemistry
Background:
- Protein crystallization is essential for X-ray structure determination but often challenging.
- Commercial screens are widely used but do not guarantee crystallization for all proteins.
- Chemical modification can enhance the crystallizability of recalcitrant proteins.
Purpose of the Study:
- To introduce a simple, cost-effective method for protein crystallization via reductive methylation.
- To assess the impact of lysine residue methylation on protein physico-chemical properties.
- To demonstrate the utility of this method for obtaining diffraction-quality crystals and structures.
Main Methods:
- Reductive methylation of lysine residues in ten different proteins.
- Analysis of changes in protein physico-chemical properties post-methylation.
- X-ray diffraction data collection and structure determination for crystallized proteins.
Main Results:
- Reductive methylation was successfully applied to ten proteins.
- The method altered protein physico-chemical properties.
- Diffraction-quality crystals were obtained for four proteins.
- X-ray structures were determined for three proteins that were previously difficult to crystallize.
Conclusions:
- Reductive methylation of lysine residues is an efficient strategy to overcome protein crystallization challenges.
- This method is applicable to a range of proteins and suitable for both low- and high-throughput settings.
- The technique facilitates the structure determination of previously intractable proteins.
