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Title: The exquisite specificity of human protein arginine methyltransferase 7 (PRMT7) toward Arg-X-Arg sites
Mammalian protein arginine methyltransferase 7 (PRMT7) has been shown to target substrates with motifs containing two arginine residues separated by one other residue (RXR motifs). In particular, the repression domain of human histone H2B (29-RKRSR-33) has been a key substrate in determining PRMT7 activity. We show that incubating human PRMT7 and [3H]-AdoMet with full-lengthXenopus laevishistone H2B, containing the substitutions K30R and R31K (RKRSR to RRKSR), results in greatly reduced methylation activity. Using synthetic peptides, we have now focused on the enzymology behind this specificity. We show for the human and Xenopus peptide sequences 23–37 the difference in activity results from changes in the Vmaxrather than the apparent binding affinity of the enzyme for the substrates. We then characterized six additional peptides containing a single arginine or a pair of arginine residues flanked by glycine and lysine residues. We have corroborated previous findings that peptides with an RXR motif have much higher activity than peptides that contain only one Arg residue. We show that these peptides have similar apparent kmvalues but significant differences in their Vmaxvalues. Finally, we have examined the effect of ionic strength on these peptides. We found the inclusion of salt had little effect on the Vmaxvalue but a considerable increase in the apparent kmvalue, suggesting that the inhibitory effect of ionic strength on PRMT7 activity occurs largely by decreasing apparent substrate-enzyme binding affinity. In summary, we find that even subtle substitutions in the RXR recognition motif can dramatically affect PRMT7 catalysis.  more » « less
Award ID(s):
1714569
PAR ID:
10476860
Author(s) / Creator(s):
; ;
Editor(s):
Ganesan, A
Publisher / Repository:
Public Library of Science
Date Published:
Journal Name:
PLOS ONE
Volume:
18
Issue:
5
ISSN:
1932-6203
Page Range / eLocation ID:
e0285812
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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