LC-MS-MS quantitative analysis reveals the association between FTO and DNA methylation

LC-MS-MS quantitative analysis reveals the association between FTO and DNA methylation

Fat mass and obesity-associated protein (FTO) is α-ketoglutarate-dependent dioxygenase and responsible for demethylating N6-methyladenosine (m6A) in mRNA, 3-methylthymine (m3T) in single-stranded DNA (ssDNA) and 3-methyluracil (m3U) in single-stranded RNA (ssRNA). Its other function remains unknown but thousands of mammalian DNA show 5-methyl-2'-deoxycytidine (5mdC) modification and 5mdC demeth...

Label-Free Quantitative Analysis Using LC/MS

Quantitative protein determination for CYP induction via LC-MS/MS.

The Cytochrome P450 (CYP) proteins are a family of membrane bound proteins that function as a major metabolizing enzyme in the human body. Quantification of CYP induction is critical in determining the disposition, safety and efficacy of drugs in humans. Described is a gel-free, high-throughput LC-MS approach to quantitate the CYP isoforms 1A2, 2B6, 3A4 and 3A5 by measuring isoform specific pep...

Chemical Analysis of Pesticides Using GC/MS, GC/MS/MS, and LC/MS/MS

There are well over 500 registered pesticides worldwide for use in agricultural regions and new agrochemicals are introduced to the marketplace continuously. This chapter deals with the chemical analysis methods for the main pesticide chemical classes that are most frequently analyzed with gas chromatography (GC) or liquid chromatography (LC) coupled to mass spectrometry (MS). GC amenable pesti...

LC/MS analysis of cellular RNA reveals NAD-linked RNA

We developed a general method to detect cellular small molecule-RNA conjugates that does not rely on the reactivity of the small molecule. This technique revealed NAD-linked RNA in Escherichia coli and Streptomyces venezuelae. Subsequent characterization showed that NAD is a 5' modification of RNA, cannot be installed in vitro through aberrant transcriptional initiation, is only found among sma...