Arginine Methylation by PRMT1 Affects ADAMTS13 Secretion and Enzymatic Activity. Arterioscler Thromb Vasc Biol 2025 Apr;45(4):506-522
Date
02/13/2025Pubmed ID
39945068Pubmed Central ID
PMC11945488DOI
10.1161/ATVBAHA.124.322249Scopus ID
2-s2.0-85219421686 (requires institutional sign-in at Scopus site) 2 CitationsAbstract
BACKGROUND: ADAMTS13 (a disintegrin and metalloprotease with thrombospondin type 1 repeats, 13), primarily synthesized in hepatic stellate and endothelial cells, plays a pivotal role in regulation of hemostasis by proteolytic cleavage of von Willebrand factor. Severe deficiency of plasma ADAMTS13 activity may result in thrombotic thrombocytopenic purpura, a potentially fatal blood disorder. ADAMTS13 undergoes posttranslational modifications including glycosylation, citrullination, oxidation. The present study determines the impact of arginine methylation by PRMT1 (protein arginine methyltransferase 1) on ADAMTS13 secretion and function.
METHODS: Cell culture, recombinant protein, biochemical analysis, site-directed mutagenesis, and animal models were utilized.
RESULTS: An inhibition of arginine methylation by a type I methyl transferase PRMT inhibitor (MS023) in HEK (human embryonic kidney) 293 cells expressing recombinant ADAMTS13 and in mice results in a significant reduction of ADAMTS13 secretion, but the secreted ADAMTS13 shows an increased specific activity; conversely, an overexpression of PRMT1 in HEK-293 cells and in transgenic mice results in an increase of ADAMTS13 secretion, but the secreted ADAMTS13 exhibits a significantly reduced specific activity. The altered ADAMTS13 activity appeared to be related to its conformational changes. LC-MS/MS (liquid chromatography with tandem mass spectrometry) identified greater than100 arginine methylation events on purified recombinant ADAMTS13. Site-directed mutagenesis performed on 5 highly conserved methylation sites (R193, R498, R692, R1123, and R1206) identifies the critical role of R1206 in ADAMTS13 function. The ADAMTS13 R1206K variant exhibits a 4- to 5-fold increase of specific activity, likely resulting from an alleviation of allosteric inhibition.
CONCLUSIONS: These results demonstrate the crucial role of arginine methylation in ADAMTS13 secretion and function. Our findings may shed new light on the mechanism of allosteric regulation of ADAMTS13, which may have a therapeutic implication.
Author List
Liu S, Ma M, Qu J, Muia J, Wu Z, Bonnez Q, Vanhoorelbeke K, Zheng L, Zhao X, Zheng XLAuthor
Joshua Muia PhD Assistant Professor in the Biochemistry department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
ADAM ProteinsADAMTS13 Protein
Animals
Arginine
Enzyme Inhibitors
HEK293 Cells
Humans
Methylation
Mice
Mice, Transgenic
Mutagenesis, Site-Directed
Protein Processing, Post-Translational
Protein-Arginine N-Methyltransferases
Repressor Proteins
Transfection
