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Highly sensitive detection of M.SssI DNA methyltransferase activity using a personal glucose meter.

Related Articles Highly sensitive detection of M.SssI DNA methyltransferase activity using a personal glucose meter. Anal Bioanal Chem. 2016 Aug;408(21):5867-72 Authors: Deng H, Peng SY, Gao Z Abstract A simple method for highly sensitive and selective detection of M.SssI CpG methyltransferase (M.SssI MTase) activity is developed, leveraging on the portability and ease of use of a personal glucose meter (PGM). Briefly, DNA-invertase conjugates are hybridized with their complementary DNA strands pre-immobilized on magnetic beads. The 5'-CCGG-3' sequence present in the DNA duplexes serves as the recognition site for both Hpa II restriction enzyme and M.SssI MTase (5'-CG-3'). Hpa II restriction enzyme specifically cleaves at unmethylated 5'-CCGG-3' sequence, and the invertase that remains on the methylated DNA catalyzes the hydrolysis of sucrose to glucose and fructose. It is found that the amount of glucose is proportional to the M.SssI MTase methylation activity in the range of 0.5 to 80 U/mL with a detection limit of 0.37 U/mL. Due to the specific recognition sequence present in the DNA strands, this method also shows high selectivity for M.SssI MTase. In addition, inhibition studies with 5'-azacytidine demonstrate the capability of inhibition screening using this method. Graphical abstract Deteciton of M.SssI DNA methyltransferase activity by a personal glucose meter. PMID: 27311957 [PubMed - indexed for MEDLINE]

Synthesis of bifunctional [email protected](OH)[email protected]@TiO2 sandwich-like nanosheets for sequential selective enrichment of phosphopeptides and glycopeptides for mass spectrometric analysis.

Related Articles Synthesis of bifunctional [email protected](OH)[email protected]@TiO2 sandwich-like nanosheets for sequential selective enrichment of phosphopeptides and glycopeptides for mass spectrometric analysis. Anal Bioanal Chem. 2016 Aug;408(20):5489-97 Authors: Xu D, Gao M, Deng C, Zhang X Abstract In this work, the bifunctional [email protected](OH)[email protected]@TiO2 sandwich-like nanosheets were designed and synthesized for the sequential selective enrichment of phosphopeptides and glycopeptides. Due to the bifunctional property of the titanium dioxide and the boronic acid group, the nanosheets were successfully applied to the enrichment of phosphopeptides and glycopeptides sequentially, evaluated by capturing phosphopeptides from tryptic digestion of model phosphoprotein bovine β-casein diluted to 0.02 ng/μL (8 × 10(-16) mol/μL) and glycopeptides from tryptic digestion of model glycoprotein horseradish peroxidase (HRP) diluted to 0.1 ng/μL (2.5 × 10(-15) mol/μL). The enrichment selectivity of the bifunctional nanosheets was evaluated by capturing phosphopeptides from a peptide mixture of β-casein and bovine serum albumin (BSA) with the molar ratio of 1:1000 (8.3 × 10(-12) mol of β-casein and 8.3 × 10(-9) mol of BSA in 100 μL) and glycopeptides from a peptide mixture of HRP and BSA up to the ratio of 1:50 (5.0 × 10(-11) mol of HRP and 2.5 × 10(-9) mol of BSA in 100 μL). Graphical Abstract A workflow of the sequential enrichment strategy for phosphopeptides and glycopeptides by the bifunctional [email protected](OH)[email protected]@TiO2 sandwich-like nanosheets. PMID: 27236315 [PubMed - indexed for MEDLINE]