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Quantitation of Staphylococcal Protein A: Determination of Equilibrium Constant and Number of Protein a Residues on Bacteria¹

From the Departments of Medicine and Pediatrics, University of Minnesota, Minneapolis, Minnesota 55455, and the Department of Medicine, University of New Mexico, Albuquerque, New Mexico 87106

Abstract

For quantitation of cell wall associated staphylococcal protein A a method was developed using ¹²⁵I-labeled myeloma globulins. The maximal sensitivity of the method could be varied according to the particular experimental questions. Using the same reagent the number of protein A residues on Staphylococcus aureus, Cowan I, was estimated to 80,000/organism. In comparison with the blood group antigen A the protein A residue is four times as densely located on this staphylococcal strain. The equilibrium constants for the reactions between two myeloma globulins and staphylococcal protein A were found to be 4.0 x 10⁷ and 4.5 x 10⁷ liters/mol, respectively.

Footnotes

This work was supported in part by United States Public Health Service Grants AM-AI-13824, AM-13690, and AI-08821.

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