Nuclear magnetic resonance spectroscopic characterization of peptide based nanocarriers and cargo complexes

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Banwo, Inumidun Damilola
University of Lethbridge. Faculty of Arts and Science
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Lethbridge, Alta. : Universtiy of Lethbridge, Department of Chemistry and Biochemistry
Cell penetrating peptides (CPPs) are short peptide 8-30 amino acids polypeptides that can deliver various molecules (nanoparticles to large fragment DNA) to subcellular locations such as the plastid organelles, cytoplasm, and nucleus etc. CPPs are characterized by containing clusters of cationic side chains that allow them to interact directly with the polar membrane surface, which enables then to enter the cell. One of the function of CPPs are to deliver cargo molecules such as DNA and RNA into cells. Most application have been developed for animal cells; however, the use of CPPs in plant cells remain a small researched field because of insufficient understanding of their mode of uptake. The present research describes firstly, the use of NMR technique to elucidate the structural properties of CPP-DNA complexation. Secondly the dynamic of the complexation. Finally, to relate the observations to their translocation mechanism. For this study, short peptides such as; arginine (R), tri-arginine (R3), nano-arginine (R9), and Tat2 (RKKRRQRRRRKKRRQRRR) were complexed with single-stranded and double-stranded DNA 5’ AGTCC 3’. The interaction between the single-stranded and double-stranded DNA 5’ AGTCC 3’ with the peptides was compared to understand how different DNA strands can influence complex formation. These complexes are used at relatively high concentrations; hence, measurements were at millimolar concentration. Therefore, for the study of DNA-CPP complexes, both non-isotopically enriched DNA and CPP samples were used. The reason for using such samples is to see if complexation can be observed using NMR spectroscopy, which will reduce the use of costly materials.
Cells -- Permeability -- Research , Plant cell membranes -- Research , Nuclear magnetic resonance spectroscopy , Biological transport -- Physiology , Peptides -- Phsyiological transport , cargo molecules , cell penetrating peptides , cellular uptake , NMR spectroscopy , plant cells , plasma membrane passage