Equilibrium Binding Constants of C3DP, A Nucleic Acid-Binding Protein Isolated from Human Serum

Abstract
The malignancy-associated [complement component C3 derived] DNA-binding protein C3DP was isolated from normal human serum and identified by dodecyl sulfate/polyacrylamide gradient gel electrophoresis and immunodiffusion. The binding of purified C3DP to DNA and RNA was investigated by density gradient centrifugation. The equilibrium binding constant K of C3DP binding to native human [spleen] DNA was determined to be K = (2.9 .+-. 0.5) .times. 105 M-1. From this value it is concluded that C3DP binds non-specifically with regard to the base sequence of the DNA. K was found to be in the same order of magnitude when measured with single-stranded or double-stranded DNA of varying G + C content, and with [mouse leukemia L5178Y cells] RNA. It varied only slightly with the pH or ionic strength of the medium, indicating that only 1 Na ion is released from DNA when 1 molecule of C3DP is bound. Ionic interactions cannot account for the stability of the C3DP-DNA complex and therefore H-bonds and hydrophobic interactions are likely to be formed.