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Three PI3P-binding LEDs bind PI3P in vitro with different affinities. The binding of the affinity-purified GST fusion proteins of several different LEDs—LED027 from Ceg19, LED035 from Lem21, and LED006 from Ceg22 or from lpg2327 (300 pmol)—to 2-fold serial dilutions of PIs (100 to 1.56 pmol) immobilized on nitrocellulose membranes was analyzed by a protein-lipid overlay assay using an anti-GST antibody.
