Bioorthogonal profiling of protein methylation using azido derivative of S-adenosyll-(L)-methionine Journal Article
| Authors: | Islam, K.; Bothwell, I.; Chen, Y.; Sengelaub, C.; Wang, R.; Deng, H.; Luo, M. |
| Article Title: | Bioorthogonal profiling of protein methylation using azido derivative of S-adenosyll-(L)-methionine |
| Abstract: | Protein methyltransferases (PMTs) play critical roles in multiple biological processes. Because PMTs often function in vivo through forming multimeric protein complexes, dissecting their activities in the native contexts is challenging but relevant. To address such a need, we envisioned a Bioorthogonal Profiling of Protein Methylation (BPPM) technology, in which a SAM analogue cofactor can be utilized by multiple rationally engineered PMTs to label substrates of the corresponding native PMTs. Here, 4-azidobut-2-enyl derivative of S-adenosyl-l-methionine (Ab-SAM) was reported as a suitable BPPM cofactor. The resultant cofactor-enzyme pairs were implemented to label specifically the substrates of closely related PMTs (e.g., EuHMT1 and EuHMT2) in a complex cellular mixture. The BPPM approach, coupled with mass spectrometric analysis, enables the identification of the nonhistone targets of EuHMT1/2. Comparison of EuHMT1/2s methylomes indicates that the two human PMTs, although similar in terms of their primary sequences, can act on the distinct sets of nonhistone targets. Given the conserved active sites of PMTs, Ab-SAM and its use in BPPM are expected to be transferable to other PMTs for target identification. © 2012 American Chemical Society. |
| Keywords: | unclassified drug; methylation; sequence analysis; mass spectrometry; proteins; proteome; reproducibility of results; nonhistone protein; protein targeting; methyltransferases; sequence homology, amino acid; in-vivo; alkylation; amino acids; protein methylation; click chemistry; coenzymes; protein methyltransferases; s-adenosylmethionine; cofactors; hek293 cells; protein methyltransferase; active site; biological process; azide; azides; protein engineering; target identification; azido derivatives; label substrates; mass spectrometric analysis; multimeric; primary sequences; protein complexes; s adenosyl l methionines; 4 azidobut 2 enylderivative; methionine derivative; s adenosyllevo methionine; bioorthogonal profiling of protein methylation |
| Journal Title: | Journal of the American Chemical Society |
| Volume: | 134 |
| Issue: | 13 |
| ISSN: | 0002-7863 |
| Publisher: | American Chemical Society |
| Date Published: | 2012-04-04 |
| Start Page: | 5909 |
| End Page: | 5915 |
| Language: | English |
| DOI: | 10.1021/ja2118333 |
| PROVIDER: | scopus |
| PMCID: | PMC3336210 |
| PUBMED: | 22404544 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 1 May 2012" - "CODEN: JACSA" - "Source: Scopus" |
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