Document Detail

Continuous enzymatic assay for histone lysine methyltransferases.
MedLine Citation:
PMID:  18072589     Owner:  NLM     Status:  MEDLINE    
We describe a continuous peptide methylation assay using the Neurospora crassa Dim-5 histone H3 lysine 9 (H3K9) methyltransferase as a model system. The assay uses streptavidin FlashPlates coated with target peptide. Since no washing and pipeting steps were required after the addition of the enzyme/S-adenosyl-L-methionine (AdoMet) mixture to the microplate, a continuous readout of the reaction progress was possible. We show that this assay is highly reproducible (with errors in the order of +/- 3%). The continuous assay is well suited for the simultaneous analysis of up to 384 samples, thus allowing for a rapid screening of methylation rates of different substrates under different conditions or in the presence of inhibitors.
Philipp Rathert; Xiaodong Cheng; Albert Jeltsch
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  BioTechniques     Volume:  43     ISSN:  0736-6205     ISO Abbreviation:  BioTechniques     Publication Date:  2007 Nov 
Date Detail:
Created Date:  2007-12-12     Completed Date:  2008-01-10     Revised Date:  2014-09-16    
Medline Journal Info:
Nlm Unique ID:  8306785     Medline TA:  Biotechniques     Country:  England    
Other Details:
Languages:  eng     Pagination:  602, 604, 606 passim     Citation Subset:  IM    
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MeSH Terms
Biological Assay / methods*
Histone-Lysine N-Methyltransferase / metabolism*
Neurospora crassa / enzymology*
Peptides / metabolism
Reproducibility of Results
S-Adenosylmethionine / metabolism
Grant Support
Reg. No./Substance:
0/Peptides; 7LP2MPO46S/S-Adenosylmethionine; EC N-Methyltransferase

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