title: Application of Celluspots peptide arrays for the analysis of the binding specificity of epigenetic reading domains to modified histone tails.
abstract: BACKGROUND
  Epigenetic reading domains are involved in the regulation of gene expression and chromatin state by interacting with histones in a post-translational modification specific manner. A detailed knowledge of the target modifications of reading domains, including enhancing and inhibiting secondary modifications, will lead to a better understanding of the biological signaling processes mediated by reading domains.
  
  
  RESULTS
  We describe the application of Celluspots peptide arrays which contain 384 histone peptides carrying 59 post translational modifications in different combinations as an inexpensive, reliable and fast method for initial screening for specific interactions of reading domains with modified histone peptides. To validate the method, we tested the binding specificities of seven known epigenetic reading domains on Celluspots peptide arrays, viz. the HP1ß and MPP8 Chromo domains, JMJD2A and 53BP1 Tudor domains, Dnmt3a PWWP domain, Rag2 PHD domain and BRD2 Bromo domain. In general, the binding results agreed with literature data with respect to the primary specificity of the reading domains, but in almost all cases we obtained additional new information concerning the influence of secondary modifications surrounding the target modification.
  
  
  CONCLUSIONS
  We conclude that Celluspots peptide arrays are powerful screening tools for studying the specificity of putative reading domains binding to modified histone peptides.
authors: Biochemistry Laboratory, School of Engineering and Science, Jacobs University Bremen, Campus Ring 1, 28759 Bremen, Germany.
authors: Ina
authors: I
authors: Bock
authors: Srikanth
authors: S
authors: Kudithipudi
authors: Raluca
authors: R
authors: Tamas
authors: Goran
authors: G
authors: Kungulovski
authors: Arunkumar
authors: A
authors: Dhayalan
authors: Albert
authors: A
authors: Jeltsch
mesh_terms: D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000378:metabolism; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000737:chemistry / Q000378:metabolism; D000090205:DNA Methyltransferase 3A; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D044127:Epigenesis, Genetic; D006657:Histones / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism; D056484:Jumonji Domain-Containing Histone Demethylases / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism*; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity; D014157:Transcription Factors; D000071857:Tumor Suppressor p53-Binding Protein 1; D000092468:Bromodomain Containing Proteins
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; D004268:DNA-Binding Proteins; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D056484:Jumonji Domain-Containing Histone Demethylases; D009687:Nuclear Proteins; D010446:Peptide Fragments; D010750:Phosphoproteins; D017346:Protein Serine-Threonine Kinases; D014157:Transcription Factors; D000071857:Tumor Suppressor p53-Binding Protein 1; C098415:BRD2 protein, human; D000092468:Bromodomain Containing Proteins; C000717967:DNMT3A protein, human; C547278:MPHOSPH8 protein, human; C499311:RAG2 protein, human; C498301:TP53BP1 protein, human; C494424:KDM4A protein, human