title: Specific and efficient N-propionylation of histones with propionic acid N-hydroxysuccinimide ester for histone marks characterization by LC-MS.
abstract: Histones participate in epigenetic regulation via a variety of dynamic posttranslational modifications (PTMs) on them. Mass spectrometry (MS) has become a powerful tool to investigate histone PTMs. With the bottom-up mass spectrometry approach, chemical derivatization of histones with propionic anhydride or deuterated acetic anhydride followed by trypsin digestion was widely used to block the hydrophilic lysine residues and generate compatible peptides for LC-MS analysis. However, certain severe side reactions (such as acylation on tyrosine or serine) caused by acid anhydrides will lead to a number of analytical issues such as reducing results accuracy and impairing the reproducibility and sensitivity of MS analysis. As an alternative approach, we report a novel derivatization method that utilizes N-hydroxysuccinimide ester to specifically and efficiently derivatize both free and monomethylated amine groups in histones. A competitive inhibiting strategy was implemented in our method to effectively prevent the side reactions. We demonstrated that our method can achieve excellent specificity and efficiency for histones derivatization in a reproducible manner. Using this derivatization method, we succeeded to quantitatively profile the histone PTMs in KMS11 cell line with selective knock out of translocated NSD2 allele (TKO) and the original parental KMS11 cell lines (PAR) (NSD2, a histone methyltransferase that catalyzes the histone H3 K36 methylation), which revealed a significant crosstalk between H3 protein K27 methylation and adjacent K36 methylation.
authors: China Novartis Institutes for BioMedical Research Co. Ltd., Building 8, lane 898 Halei road, Shanghai 201203, China.
authors: Rijing
authors: R
authors: Liao
authors: Haiping
authors: H
authors: Wu
authors: Haibing
authors: H
authors: Deng
authors: Yanyan
authors: Y
authors: Yu
authors: Min
authors: M
authors: Hu
authors: Huili
authors: H
authors: Zhai
authors: Pengyuan
authors: P
authors: Yang
authors: Shaolian
authors: S
authors: Zhou
authors: Wei
authors: W
authors: Yi
mesh_terms: D000595:Amino Acid Sequence; D000641:Ammonia / Q000737:chemistry; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008745:Methylation; D010455:Peptides / Q000032:analysis / Q000378:metabolism; D011422:Propionates / Q000737:chemistry*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013388:Succinimides / Q000737:chemistry*; D014357:Trypsin / Q000378:metabolism
chemical_list: D006657:Histones; D010455:Peptides; D011422:Propionates; D012097:Repressor Proteins; D013388:Succinimides; D000641:Ammonia; D011495:Histone-Lysine N-Methyltransferase; C419586:NSD2 protein, human; D014357:Trypsin; C029658:propionic acid; C001426:N-hydroxysuccinimide