Quantitative assessment of the association of the α-I fragment of spectrin with oligomers of intact spectrin

Abstract

• 1.

  1. The α-I fragment of human spectrin that carries the binding site on the α-chain of spectrin for the β -chain has been purified from limited trypsin digests of spectrin by means of FPLC.

• 2.

  2. The self-association of spectrin and the binding of the α-I fragment to spectrin heterodimers and to tetramers have been quantified through the use of gel electrophoresis, staining with Coomassie Blue, and quantification of the bound dye following elution with pyridine.

• 3.

  3. The parameters of self-association were found to be consistent with those estimated from sedimentation equilibrium analysis.

• 4.

  4. The data were consistent with a model in which both self-association and the binding of the α-I fragment are considered to occur through an intermediate in which the α-β interface is initially dissociated. The α-β interface in the heterodimer was found to be less stable than that of higher oligomers by approx. 3 kJ/mol.