Affinity retardation chromatography: characterization of the method and its application. The description of low affinity laminin self-interactions.
Affinity retardation chromatography (ARC), a method for the examination of low-affinity interactions, is mathematically described in order to characterize the method itself and to estimate binding coefficients of self-assembly domains of basement membrane protein laminin. Affinity retardation was determined by comparing the elutions on a "binding" and on a "nonreacting" column. It depends on the binding coefficient, the concentrations of both ligands, and the nonbinding elution position. Half maximal binding of the NH2-terminal domain of laminin B1-short arm to the A- and/or B2-short arms was estimated to occur at 10-17 microM for noncooperative and at < or = 3 microM for cooperative binding. A model of the laminin polymerization, postulating two levels of cooperative binding behavior, is described.
Schittny
J C
JC
Department of Developmental Biology, University of Bern, Switzerland.
D002846:Chromatography, Affinity / Q000379:methods*; D007797:Laminin / Q000737:chemistry*; D008956:Models, Chemical; D011485:Protein Binding
D007797:Laminin