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. 1985 Sep 25;13(18):6621–6634. doi: 10.1093/nar/13.18.6621

Binding studies of SV40 T-antigen to SV40 binding site II.

P Gottlieb, M S Nasoff, E F Fisher, A M Walsh, M H Caruthers
PMCID: PMC321981  PMID: 2997717

Abstract

SV40 T-Antigen binding site II was synthesized, cloned and analyzed for its ability to bind purified SV40 T-antigen. We report the binding constant of T-antigen for isolated site II. Using a filter binding assay the calculated binding constant was 6-8 fold less efficient than site I previously reported. Binding constants were calculated using two methods. The first was a direct calculation using a protein titration curve (KD). The second was by the ratio of measured association and dissociation rates. Both methods gave similar constants. Protection studies with SV40 T-antigen on the T-antigen binding sites in the wild-type array demonstrated that the binding constants of site I and site II are similar to those calculated for the individual sites. These results demonstrate that SV40 T-antigen does not bind cooperatively to sites one and two as earlier believed and are in agreement with recent observations emanating from several laboratories.

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Selected References

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