1 research outputs found
Biophysical Characterization of Human Protamine‑1 as a Responsive CEST MR Contrast Agent
The protamines are a low-molecular-weight,
arginine-rich family
of nuclear proteins that protect chromosomal DNA in germ cells by
packing it densely using electrostatic interactions. Human protamine-1
(hPRM1) has been developed as a magnetic resonance imaging (MRI) chemical
exchange saturation transfer (CEST) reporter gene, based on a sequence
that is approximately 50% arginine, which has a side chain with rapidly
exchanging protons. In this study, we have synthesized hPRM1 and determined
how its CEST MRI contrast varies as a function of pH, phosphorylation
state, and upon noncovalent interaction with nucleic acids and heparin
(as antagonist). CEST contrast was found to be highly sensitive to
phosphorylation on serine residues, intra- and intermolecular disulfide
bridge formation, and the binding of negatively charged nucleotides
and heparin. In addition, the nucleotide binding constants (<i>K</i><sub>eq</sub>) for the protamines were determined through
plotting the molar concentration of heparin versus CEST contrast and
compared between hPRM1 and salmon protamine. Taken together, these
findings are important for explaining the CEST contrast of existing
arginine-rich probes as well as serving as a guideline for designing
new genetic or synthetic probes