title: Investigation of binding site density: effects on the interaction between cibacron blue-dextran conjugates and lysozyme.
abstract: Cibacron-blue-dextran conjugates have been produced with a range of ligand loadings using a dextran preparation of average molecular weight of 2 x 10(6). The equilibrium binding capacity of these ligand conjugates for lysozyme was determined using a gel permeation procedure to separate bound from free protein. The results obtained give clear evidence for at least two types of binding showing a marked difference in affinity. For the higher-affinity interaction the half-saturation constant decreases with increasing ligand loading. The number of dye molecules participating in binding is proportional to loading up to 154 mol dye/mol dextran but is reduced at the highest loading used (315 mol dye/mol dextran). This may be due to steric interference or to dye stacking reducing the number of dye molecules available for binding.
authors.affiliation: Biochemistry Department, University of Bath, Bath, BA2 7AY, United Kingdom.
authors.forename: A G
authors.initials: AG
authors.lastname: Mayes
authors.forename: J D
authors.initials: JD
authors.lastname: Moore
authors.forename: R
authors.initials: R
authors.lastname: Eisenthal
authors.forename: J
authors.initials: J
authors.lastname: Hubble