Conformational transition of giant DNA in a confined space surrounded by a phospholipid membrane.
It has been established that a long DNA molecule exhibits a large discrete conformational change from a coiled state to a highly folded state in aqueous solution, depending on the presence of various condensing agents such as polyamines. In this study, T4 DNA labeled with fluorescent dyes was encapsulated in a cell-sized microdroplet covered with a phospholipid membrane to investigate the conformational behavior of a DNA molecule in such a confined space. Fluorescence microscopy showed that the presence of Mg(2+) induced the adsorption of DNA onto the membrane inner-surface of a droplet composed of phosphatidylethanolamine, while no adsorption was observed onto a phosphatidylcholine membrane. Under the presence of spermine (tetravalent amine), DNA had a folded conformation in the bulk solution. However, when these molecules were encapsulated in the microdroplet, DNA adsorbed onto the membrane surface accompanied by unfolding of its structure into an extended coil conformation under high concentrations of Mg(2+). In addition, DNA molecules trapped in large droplets tended not to be adsorbed on the membrane, i.e., no conformational transition occurred. A thermodynamic analysis suggests that the translational entropy loss of a DNA molecule that is accompanied by adsorption is a key factor in these phenomena under micrometer-scale confinement.
Kato
Ayako
A
Graduate School of Pharmaceutical Sciences, Josai University, Sakado, Japan.
Shindo
Eri
E
Sakaue
Takahiro
T
Tsuji
Akihiko
A
Yoshikawa
Kenichi
K
D000327:Adsorption / Q000187:drug effects; D017122:Bacteriophage T4; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D019277:Entropy; D005456:Fluorescent Dyes / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D008567:Membranes, Artificial; D008856:Microscopy, Fluorescence; D009690:Nucleic Acid Conformation / Q000187:drug effects*; D009821:Oils / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000494:pharmacology; D010743:Phospholipids / Q000737:chemistry / Q000494:pharmacology*; D013096:Spermine / Q000494:pharmacology; D013499:Surface Properties; D014867:Water / Q000737:chemistry
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D005456:Fluorescent Dyes; D008567:Membranes, Artificial; D009821:Oils; D010714:Phosphatidylethanolamines; D010743:Phospholipids; D014867:Water; D013096:Spermine; D004247:DNA; D008274:Magnesium