Profiling substrates of protein arginine N-methyltransferase 3 with S-adenosyl-L-methionine analogues Journal Article
| Authors: | Guo, H.; Wang, R.; Zheng, W.; Chen, Y.; Blum, G.; Deng, H.; Luo, M. |
| Article Title: | Profiling substrates of protein arginine N-methyltransferase 3 with S-adenosyl-L-methionine analogues |
| Abstract: | Protein arginine N-methyltransferase 3 (PRMT3) belongs to the family of type I PRMTs and harbors the activity to use S-adenosyl-L-methionine (SAM) as a methyl-donor cofactor for protein arginine labeling. However, PRMT3's functions remain elusive with the lacked knowledge of its target scope in cellular settings. Inspired by the emerging Bioorthogonal Profiling of Protein Methylation (BPPM) using engineered methyltransferases and SAM analogues for target identification, the current work documents the endeavor to systematically explore the SAM-binding pocket of PRMT3 and identify suitable PRMT3 variants for BPPM. The M233G single point mutation transforms PRMT3 into a promiscuous alkyltransferase using sp2-β-sulfonium-containing SAM analogues as cofactor surrogates. Here the conserved methionine was defined as a hot spot that can be engineered alone or in combination with nearby residues to render cofactor promiscuity of multiple type I PRMTs. With this promiscuous variant and the matched 4-propargyloxy-but-2-enyl (Pob)-SAM analogue as the BPPM reagents, more than 80 novel proteins were readily uncovered as potential targets of PRMT3 in the cellular context. Subsequent target validation and functional analysis correlated the PRMT3 methylation to several biological processes such as cytoskeleton dynamics, whose roles might be compensated by other PRMTs. These BPPM-revealed substrates are primarily localized but not restricted in cytoplasm, the preferred site of PRMT3. The broad localization pattern may implicate the diverse roles of PRMT3 in the cellular setting. The revelation of PRMT3 targets and the transformative character of BPPM for other PRMTs present unprecedented pathways toward elucidating physiological and pathological roles of diverse PRMTs. © 2013 American Chemical Society. |
| Keywords: | controlled study; unclassified drug; methylation; cytoplasm; point mutation; cytoskeleton; methionine; protein arginine methyltransferase; chemical binding; s adenosylmethionine; enzyme engineering; human; priority journal; article; protein arginine methyltransferase 3 |
| Journal Title: | ACS Chemical Biology |
| Volume: | 9 |
| Issue: | 2 |
| ISSN: | 1554-8929 |
| Publisher: | American Chemical Society |
| Date Published: | 2014-02-21 |
| Start Page: | 476 |
| End Page: | 484 |
| Language: | English |
| DOI: | 10.1021/cb4008259 |
| PROVIDER: | scopus |
| PMCID: | PMC3944066 |
| PUBMED: | 24320160 |
| DOI/URL: | |
| Notes: | Export Date: 2 April 2014 -- CODEN: ACBCC -- Source: Scopus |
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