JBS Methylation Kit

Surface engineering of proteins can be a powerful technique for dealing with proteins that yield no or poorly diffracting crystals. In particularly, reductive methylation of proteins has emerged as a standard procedure in several large scale facilities and research programs, i.e. the Midwest Centre of Structural genomics [1] and the Structural Proteomics In Europe (SPINE) program [2,3].

The JBS Methylation Kit is designed for selective methylation of lysine residues. The method does not require laborious cloning/expression/purification but chemically replaces the protons of the amino group of all lysine residues with methyl groups. The result is a surface-engineered protein within 24 hours ready for crystallization.

Each JBScreen Methylation Kit contains all necessary reagents for six methylation experiments. All components are provided ready for use. Just follow the manual step-by-step. No background in chemistry necessary.

References

[1] Kim et al. (2008) Large-scale evaluation of protein reductive methylation for improving protein crystallization. Nature Methods 5:853.
[2] Fogg et al. (2006) Application of the use of high-throughput technologies to the determination of protein structures of bacterial and viral pathogens. Acta Cryst. D 62:1196.
[3] Walter et al. (2006) Lysine methylation as a routine rescue strategy for protein crystallization. Structure 14:1617.

Selected Literature Citations of JBS Methylation Kit

Fu et al. (2017) The natural product carolacton inhibits folate-dependent C1 metabolism by targeting FolD/MTHFD. Nat. Commun. 8:1529.
Barden et al. (2013) A Helical RGD Motif Promoting Cell Adhesion: Crystal Structures of the Helicobacter pylori Type IV Secretion System Pilus Protein CagL. Structure 21:1931.
Peat et al. (2013) Cyanuric acid hydrolase: evolutionary innovation by structural concatenation. Molecular Microbiology 88:1149.
Koval et al. (2013) Plant multifunctional nuclease TBN1 with unexpected phospholipase activity: structural study and reaction-mechanism analysis. Acta Cryst. D 69:213.
Reeks et al. (2013) Structure of a dimeric crenarchaeal Cas6 enzyme with an atypical active site for CRISPR RNA processing. Biochem. J. 452 (2):223.
Cima et al. (2012) Insight on an Arginine Synthesis Metabolon from the Tetrameric Structure of Yeast Acetylglutamate Kinase. PLOS one 7:e34734.