Use of Cibracon Blue-Sepharose 6B to Detect Modifications of the Non-allosteric 6-Phosphofructokinase from Escherichia Coli K-12

Abstract

In crude cell-free extracts of aerobically grown E. coli K-12, the non-allosteric form of 6-phosphofructokinase has a tetrameric molecular weight of 140000 with a low affinity � 5%) for the blue dextran chromophore-Cibracon Blue. The allosteric form has the same tetrameric molecular weight, but possesses a strong affinity for the blue dextran chromophore.

Monolithic media for applications in affinity chromatography

“…Interactions of proteins with their binding partners play a major role in various cellular events, such as signal transduction and cell proliferation. Of course, biomolecules do not exclusively interact with other biomolecules, but they also undergo highly specific binding to metal ions or artificial ligands, such as small‐molecule drugs 14–16. The underlying molecular recognition mechanisms are highly selective and as such, serve as basis for the various affinity‐based chromatographic methods.…”

“…Originally, particles mainly prepared from agarose or silica were used for AC 15, 20, 21, but since the emergence of monolithic stationary phases (MSPs) with their advantageous properties, such as rapid mass transfer, high porosity, and low backpressure at high flow rates, these materials are becoming more and more popular 7, 22, 23. Also, miniaturization is easily achieved when MSPs are chosen as supports for AC 24, 25.…”