Document Detail


Continuous enzymatic assay for histone lysine methyltransferases.
MedLine Citation:
PMID:  18072589     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Philipp Rathert; Xiaodong Cheng; Albert Jeltsch
Related Documents :
18778699 - Performance evaluation of a particle-enhanced turbidimetric cystatin c assay on the hit...
12627289 - Usefulness of western blot in serological follow-up of newborns suspected of congenital...
22980839 - Separation, purification and quantification of verbascoside from penstemon barbatus (ca...
16466649 - A two-center international evaluation of the immulite 2000 automated serum gastrin assay.
6697489 - Automated kinetic method for d-3-hydroxybutyrate in plasma or serum.
6897249 - Reversed-phase high-performance liquid chromatographic separation of saccharin, caffein...
15859639 - A fast and simple high-performance liquid chromatography/mass spectrometry method for s...
19468319 - Mycotoxin detection in human samples from patients exposed to environmental molds.
21930439 - Development of an off-line capillary column imac phosphopeptide enrichment method for l...
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    
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Biological Assay / methods*
Histone-Lysine N-Methyltransferase / metabolism*
Kinetics
Methylation
Neurospora crassa / enzymology*
Peptides / metabolism
Reproducibility of Results
S-Adenosylmethionine / metabolism
Grant Support
ID/Acronym/Agency:
GM068680/GM/NIGMS NIH HHS; R01 GM068680/GM/NIGMS NIH HHS; R01 GM068680-02/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Peptides; 7LP2MPO46S/S-Adenosylmethionine; EC 2.1.1.43/Histone-Lysine N-Methyltransferase
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Cognate putative nuclear localization signal effects strong nuclear localization of a GFP reporter a...
Next Document:  Method for isolation of PCR-ready genomic DNA from zebrafish tissues.