Stabilizing and destabilizing effects of arginine on deoxyribonucleic acid.
Aqueous arginine solution now finds a wide range of applications in biotechnology fields, including protein refolding, chromatography and virus inactivation. While progress has been made for mechanistic understanding of the effects of arginine on proteins, we have little understanding on how arginine inactivates viruses. One of the viral components is nucleic acid. We have examined the effects of arginine on the structure and thermal stability of calf thymus deoxyribonucleic acid (DNA) using circular dichroism (CD). Both NaCl and arginine reduced CD intensity. At low concentrations, arginine showed a stronger effect on CD intensity than NaCl. Both NaCl and arginine sharply increased the melting temperature at low concentrations (below 0.25 M). However, they had an opposite effect at higher concentrations. Above this concentration, NaCl gradually increased the melting temperature, leading to the onset melting temperature above 90 degrees C. On the other hand, the thermal stability in the presence of arginine reached a maximum at 0.2-0.5 M, after which further addition of arginine caused decreased melting temperature. It is most likely that the increased melting temperature at low concentration is due to electrostatic stabilization of DNA structure by both NaCl and arginine and that the opposite effects at higher salt concentration are due to salt-specific effects, i.e., stabilizing (salting-out) effects of NaCl and destabilizing (salting-in) effects of arginine. Solubility measurements of nucleic acid bases showed that arginine, but not NaCl, increases the solubilities of the bases, supporting their effects on DNA stability at higher concentration.
Arakawa
Tsutomu
T
Alliance Protein Laboratories, 3957 Corte Cancion, Thousand Oaks, CA 91360, USA. tarakawa2@aol.com
Hirano
Atsushi
A
Shiraki
Kentaro
K
Kita
Yoshiko
Y
Koyama
A Hajime
AH
D001120:Arginine / Q000494:pharmacology*; D002942:Circular Dichroism; D004247:DNA / Q000187:drug effects*; D009691:Nucleic Acid Denaturation / Q000187:drug effects; D012965:Sodium Chloride / Q000494:pharmacology; D012995:Solubility / Q000187:drug effects; D013696:Temperature
D012965:Sodium Chloride; D004247:DNA; D001120:Arginine