Cooperativity in the binding of the cationic biocide polyhexamethylene biguanide to nucleic acids.
The interaction between the broad-spectrum antimicrobial agent, polyhexamethylene biguanide (PHMB), and various nucleic acids was investigated. Titration of either single- or double-stranded 100-bp DNA, or mixed-molecular weight marker DNA, or tRNA with PHMB caused precipitation of a complex between nucleic acid and PHMB in which the nucleotide/biguanide ratio was always close to unity. Binding of PHMB was highly cooperative, with apparent Hill coefficients 10.3-14.6. When a fluorescent derivative of PHMB was titrated with increasing amounts of nucleic acid, all four forms of nucleic acid caused strong polarisation of fluorescence, demonstrating the association with PHMB. The intensity and broad-spectrum binding of PHMB to all forms of nucleic acid has significant implications for the mechanism of action of this biocide.
Allen
Michael J
MJ
Cardiff School of Biosciences, Cardiff University, Cardiff CF10 3US, UK.
Morby
Andrew P
AP
White
Graham F
GF
D000890:Anti-Infective Agents / Q000737:chemistry / Q000378:metabolism; D001645:Biguanides / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002412:Cations; D011232:Chemical Precipitation; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005454:Fluorescence Polarization; D007700:Kinetics; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism*; D017186:Titrimetry / Q000379:methods
D000890:Anti-Infective Agents; D001645:Biguanides; D002412:Cations; C031233:polihexanide; D004247:DNA; D012343:RNA, Transfer