Artificial cell-spreading of permeabilized BHK cells on fibronectin-coated substratum.
We developed a cell-spreading model of permeabilized BHK cells to analyze molecular interactions in the cell-spreading machinery. BHK cells in suspension were allowed to attach onto the fibronectin-coated cover slips at 37 degrees C for 20 min. The cells were then permeabilized with 50 micrograms/ml saponin for 10 min at 4 degrees C. Approximately 50 percent of the cell models spread in response to exogenous 1 mM ATP for 90 min at 37 degrees C. Anti-actin antibody penetrated into almost all the cell models which spread ATP-dependently. These indicate that half number of the cell models permeabilized with saponin is spread by the exogenous ATP and has pores larger than an IgG molecule.
Hayashi
M
M
Department of Biology, Ochanomisu University, Tokyo, Japan.
Hamano
T
T
Ishikawa-Sakurai
M
M
Miyamoto
Y
Y
D000255:Adenosine Triphosphate / Q000493:pharmacokinetics*; D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line / Q000166:cytology; D002463:Cell Membrane Permeability / Q000502:physiology*; D048429:Cell Size / Q000502:physiology; D002592:Cetomacrogol / Q000494:pharmacology; D006224:Cricetinae; D004072:Digitonin / Q000494:pharmacology; D005353:Fibronectins / Q000502:physiology*; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D007668:Kidney / Q000166:cytology*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D012503:Saponins / Q000494:pharmacology
D005353:Fibronectins; D007074:Immunoglobulin G; D009842:Oligopeptides; D012503:Saponins; C047981:arginyl-glycyl-aspartic acid; D000255:Adenosine Triphosphate; D002592:Cetomacrogol; D004072:Digitonin