“CTP” cinnamyltriphenylphosphonium chloride Cytotoxin Molecular Weight: 414.906221 g/mol Structure: Three benzene rings and cinnamyl group attached to phosphorous central atom. Chloride anion. Physical Properties: Melting Point: 225 º – 227 º C Appearance: White to Off-White Powder or Crystalline Powder

IR Spectrum of CTP If a  is being absorbed, it means that energy (E = h) is being absorbed by the cmpd. Energies in the IR region correspond to the energies involved in bond vibrations (stretches/bends). 3100 - 3000 =C-H (aryl) stretch 3000 - 2850 C-H (alkyl) stretch 2000 - 1665 weak overtones of aromatic 1470 - 1450 C-H (alkyl) bend 1600 - 1585, 1500 -1400 C=C stretch (in ring) 1000 - 650 =C-H (alkene) bend

DNA Melting Curve dsDNA 2 ssDNA = Melting Point of DNA Half of DNA has uncoiled and remains single-stranded when the melting temperature is reached Tm A shift in Melting Temperature is a strong indicator of the presence of an interaction between the DNA and any small molecule, such as any arylphosphonium salt. Sample Native Salmon DNA Salmon DNA + 3µL (15.2µg/150µL DMSO) CTP Salmon DNA + 5µL Melting Temp. (ºC) 64.6 67.2 67.8

Gel Electrophoresis Lane 2: (Control) Digested DNA Lane 3: (Experimental) Digested DNA w/ 1ml CTP

HyperChem Predictions CTP would bind effectively with the DNA through intercalation. This was based upon the flat, characteristic structure of the cinnamyl group. CTP might also bind with DNA in its major groove, based on the relative size of the benzene groups and the electrostatic attraction of the phosphorous backbone of DNA.

HyperChem Computations Minor groove interactions were most favorable with cinnamyl group in closest proximity to DNA: Distance of two closest atoms = 3.8913 Å E = 14.699580 kcal Gd = 0.099771

HyperChem Predictions

HyperChem Computations Intercalation and Major Groove Interactions of CTP and DNA were of greater energy: Major Groove - Triphenyl group of closest proximity to DNA E = 894.84832 kcal Cinnamyl group E = 898.00171 kcal Intercalation – (Interaction optimized with CTP passing through nitrogen base of cytosine) E = 1553.85107 kcal Cinnamyl group through base pairs in major groove E = 923.01587 kcal

Further Research Nature of DNA Cooling after denaturation and possible restructuring of DNA DNA Structure effects on Movement through gel Potential Applications of the CTP Molecule in DNA of organisms Further HyperChem Computations Ultimate objective: X-ray crystallography (orientation and type of interaction)