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Journal of Biochemistry Advance Access published online on May 7, 2008
Journal of Biochemistry, doi:10.1093/jb/mvn061
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© 2008 The Japanese Biochemical Society

Protein Methyltransferase Activities in Commercial In vitro Translation Systems

Robert B. Denman*

*Direct correspondence to: Dr. Robert B. Denman. Head Biochemical Molecular Neurobiology Laboratory, Department of Molecular Biology, New York State Institute for Basic Research, In Developmental Disabilities, 1050 Forest Hill Road, Staten Island, NY 10314. Tel: (718) 494-5199, Fax: (718) 494-5905, e-mail: rbdenman{at}yahoo.com

Received April 18, 2008; Accepted April 25, 2008


  Abstract

Summary

Protein arginine methylation is a well-known post-translational modification that has been shown to occur in rabbit reticulocyte in vitro translation lysates (RRL); however, it is not known whether this is a general feature of in vitro-produced proteins from other eukaryotic cell-free translation systems, particularly insect-derived lysates (ICL). Because methylation can affect protein localization, RNA binding and protein-protein interactions this may be of great importance as in vitro-produced proteins are often used in assays of protein function. Here, I report the presence of base-stable and base-labile methyltransferase activities in RRL, ICL and wheat germ in vitro extracts (WGE). Indeed, the presence of CARM1 in RRL and ICL and a class II protein arginine methyltransferase activity (PRMT5/7) is documented in all three systems. Additionally, the lysine methyltransferase that modifies eukaryotic elongation factor 1A (eEF-1A) was detected in ICL and WGE. Importantly, using a defined set of substrates under identical conditions I show that all three in vitro systems contain different complements of the various methyltransferases. These data suggest that three systems can be used in a complementary fashion to investigate the effect(s) of post-translational modification on protein function.

Key Words: fragile x mental retardation protein, histone, myelin basic protein, protein arginine methyltransferase, protein lysine methyltransferase


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