- Selective Reagent for Detection of N-ε-Monomethylation of a Peptide Lysine Residue through SNAr Reaction
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Methylations of specific lysine residues of histone proteins are catalyzed by histone methyltransferases (HMTs) and play key roles in the epigenetic control of gene expression. Several methods to detect N-ε-methylation of the lysine residue have been established in order to evaluate the activity of HMTs, to develop inhibitors, and to identify substrates. However, they mostly employ specific antibodies or enzymes such as peptidases, and their reliability and reproducibility often depend on the quality of the protein reagents and the reaction conditions. Here, we describe a convenient method to detect N-ε-monomethylation of the lysine residue through a simple chemical reaction. We focused on nucleophilic aromatic substitution reaction (SNAr reaction) between an aromatic electrophile and a primary or monomethylated amino group. Screening of various electrophiles indicated that 4-fluoro-2-nitroacetophenone (1g) has high selectivity for the N-ε-monomethylated amino group of lysine. Furthermore, the reaction products of 1g with lysine and N-ε-monomethylated lysine, 5g and 6g, respectively, show different absorption spectra, that is, the absorbance at 350 nm of 6g is 13 times larger than that of 5g. We show that these characteristic properties of 1g can be utilized for the selective detection of the methylation state of lysine residues in HMT substrate peptides, and for an assay of HMT activity.
- Mori, Shuichi,Hirano, Tomoya,Takaguchi, Asuka,Fujiwara, Takashi,Okazaki, Yusuke,Kagechika, Hiroyuki
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supporting information
p. 3606 - 3611
(2017/07/22)
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- Compounds and Compositions as Channel Activating Protease Inhibitors
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The invention provides compounds and pharmaceutical compositions thereof, which are useful for modulating channel activating proteases, and methods for, using such compounds to treat, ameliorate or prevent a condition associated with a channel activating protease, including but not limited to prostasin, PRSS22, TMPRSS11 (e.g., TMPRSS11B, TMPRSS11E), TMPRSS2, TMPRSS3, TMPRSS4 (MTSP-2), matriptase (MTSP-1), CAP2, CAP3, trypsin, cathepsin A, or neutrophil elastase.
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Page/Page column 30-31
(2008/06/13)
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- Subtype selective substrates for histone deacetylases
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To probe the steric requirements for deacylation, we synthesized lysine-derived small molecule substrates and examined structure-reactivity relationships with various histone deacetylases. Rat liver, human HeLa, and human recombinant class I and II histon
- Heltweg, Birgit,Dequiedt, Franck,Marshall, Brett L.,Brauch, Carsten,Yoshida, Minoru,Nishino, Norikazu,Verdin, Eric,Jung, Manfred
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p. 5235 - 5243
(2007/10/03)
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- Design, synthesis, and structure-activity relationships of macrocyclic hydroxamic acids that inhibit tumor necrosis factor α release in vitro and in vivo
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To search for TNF-α (tumor necrosis factor α) converting enzyme (TACE) inhibitors, we designed a new class of macrocyclic hydroxamic acids by linking the P1 and P2′ residues of acyclic anti-succinate-based hydroxamic acids. A variety of residues including amide, carbamate, alkyl, sulfonamido, Boc-amino, and amino were found to be suitable P1 P1-P2′ linkers. With an N-methylamide at P3′, the 13-16-membered macrocycles prepared exhibited low micromolar activities in the inhibition of TNF-α release from LPS-stimulated human whole blood. Further elaboration in the P3′-P4′ area using the cyclophane and cyclic carbamate templates led to the identification of a number of potent analogues with IC50 values of ≤0.2 μM in whole blood assay (WBA). Although the P3′ area can accommodate a broad array of structurally diversified functional groups including polar residues, hydrophobic residues, and amino and carboxylic acid moieties, in both the cyclophane series and the cyclic carbamate series, a glycine residue at P3′ was identified as a critical structural component to achieve both good in vitro potency and good oral activity. With a glycine residue at P3′, an N-methylamide at P4′ provided the best cyclophane analogue, SL422 (WBA IC50 = 0.22 μM, LPS-mouse ED50 = 15 mg/kg, po), whereas a morpholinylamide at P4′ afforded the most potent and most orally active cyclic carbamate analogue, SP057 (WBAIC50 = 0.067 μM, LPS-mouse ED50 = 2.3 mg/kg, po). Further profiling for SL422 and SP057 showed that these macrocyclic compounds are potent TACE inhibitors, with Ki values of 12 and 4.2 nM in the porcine TACE assay, and are broad-spectrum MMP inhibitors. Pharmacokinetic studies in beagle dogs revealed that SL422 and SP057 are orally bioavailable, with oral bioavailabilities of 11% and 23%, respectively.
- Xue,Voss,Nelson,Duan,Cherney,Jacobson,He,Roderick,Chen,Corbett,Wang,Meyer,Kennedy,Degrado,Hardman,Teleha,Jaffee,Liu,Copeland,Covington,Christ,Trzaskos,Newton,Magolda,Wexler,Decicco
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p. 2636 - 2660
(2007/10/03)
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- Oligopeptides derived from C-reactive protein fragments
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PCT No. PCT/EP94/01574 Sec. 371 Date Jun. 11, 1996 Sec. 102(e) Date Jun. 11, 1996 PCT Filed May 16, 1994 PCT Pub. No. WO95/10531 PCT Pub. Date Apr. 20, 1995Oligopeptides derived from fragments of C-reactive proteins and their use as immunomodulating agent
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