posted on 2019-04-29, 00:00authored byAbhishek Thakur, Joan M. Hevel, Orlando Acevedo
Protein
arginine methyltransferase 7 (PRMT7) catalyzes the formation
of monomethylarginine (MMA) but is incapable of performing a
dimethylation. Given that PRMT7 performs vital functions in mammalian
cells and has been implicated in a variety of diseases, including
breast cancer and age-related obesity, elucidating the origin of its
strict monomethylation activity is of considerable interest. Three
active site residues, Glu172, Phe71, and Gln329, have been reported
as particularly important for product specificity and enzymatic activity.
To better understand their roles, mixed quantum and molecular mechanical
(QM/MM) calculations coupled to molecular dynamics and free energy
perturbation theory were carried out for the WT, F71I, and Q329S trypanosomal
PRMT7 (TbPRMT7) enzymes bound with S-adenosyl-L-methionine (AdoMet) and an arginine substrate in an unmethylated
or methylated form. The Q329S mutation, which experimentally abolished
enzymatic activity, was appropriately computed to give an outsized
ΔG‡ of 30.1 kcal/mol for
MMA formation compared to 16.9 kcal/mol for WT. The F71I mutation,
which has been experimentally shown to convert the enzyme from a type
III PRMT into a mixed type I/II capable of forming dimethylated arginine
products, yielded a reasonable ΔG‡ of 21.9 kcal/mol for the second turnover compared to 28.8 kcal/mol
in the WT enzyme. Similar active site orientations for both WT and
F71I TbPRMT7 allowed Glu172 and Gln329 to better orient the substrate
for SN2 methylation, enhanced the nucleophilicity of the
attacking guanidino group by reducing positive charge, and facilitated
the binding of the subsequent methylated products.