Histone Modification Analysis Using Mass Spectrometry

Histone Modification Analysis Using Mass Spectrometry 

Histones are very basic proteins with an isoelectric point between 10.31 and 11.27 for human complement. They are present in virtually all eukaryotes (with the exception of dinoflagellates [14]) where they are associated with most of the nuclear DNA. The DNA is wrapped around an octamer formed by the four core histones H2A, H2B, H3 and H4 to build a nucleosome. This particle is the fundamental repeating unit of chromatin [15]. A string of nudeosomes can fold into a higher order structure, the exact molecular nature of which is still not fully understood but clearly has a strong influence on gene expression. 

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Histone Modification Analysis Using Mass Spectrometry... 

Mass spectrometry provides a more direct and precise technique to study histone modifications. As with the other methods discussed above, mass spectrometry also has several pitfalls that should be taken into account when analyzing histone modifications. First of all histones and especially the core histones H3 and H4 are rich in lysine residues. Consequently, trypsin as an enzyme that is routinely used for the identification of proteins via peptide mass fingerprints cannot be used for regular in gel digestion of histones. Other enzymes that have a different specificity (such as Asp-N or Arg-C) are more frequently used in the analysis of histones [25]. A drawback... 

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