Electromagnetophoretic force measurement of a single binding interaction between lectin and yeast cell surfaces.
A novel measurement method of the binding force between a micrometer-sized particle and a solid surface in an electrolyte solution has been established by using the electromagnetophoretic buoyancy on the particle. By this method, we investigated the binding force between a yeast cell surface and an oligosaccharide-binding protein, concanavalin A (Con A), fixed on a silica capillary wall. The force measurement was carried out up to 60 pN. In a lower surface concentration of Con A, yeast cells could be desorbed by a force less than 60 pN. However, in a higher surface concentration after treated by 1 mg ml(-1) solution, yeast cells were adsorbed with a force stronger than 60 pN. In this case, the addition of 10 mg ml(-1) D-mannose solution to the medium reduced the binding force to less than 60 pN. The observed adsorption force of yeast cells ranged within 30 - 40 pN, regardless of the interfacial amount of Con A. This force was thought to be the single binding force between a mannose group of the cell surface and an active site of Con A. Moreover, the dissociation rate constant of the single binding of yeast cell and Con A complex was determined as 4.6 x 10(-3) s(-1) and the increment of the binding distance at the transition state as 0.33 nm from the desorption kinetic experiments of yeast cell under the constant pulling conditions of 10, 20 and 30 pN. Such satisfactory results demonstrate the novel advantages of the present method.
Iiguni
Yoshinori
Y
Department of Chemistry, Graduate School of Science, Osaka University, Japan.
Watarai
Hitoshi
H
D000465:Algorithms; D003208:Concanavalin A / Q000737:chemistry*; D004574:Electromagnetic Fields; D019075:Electrophoresis, Capillary; D006860:Hydrogen Bonding; D007700:Kinetics; D037102:Lectins / Q000737:chemistry*; D008351:Mannans / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D012822:Silicon Dioxide; D013050:Spectrometry, Fluorescence
D037102:Lectins; D008351:Mannans; D011134:Polysaccharides; D003208:Concanavalin A; D012822:Silicon Dioxide