Figure 2.
Ca2+ coordination by Glu465 and Glu468 is critical to structural integrity of the sTFR. Purified sTFR (100 ng, lane 1) served as a standard. Following purification, aliquots from tissue culture batches 4, 5 and 6 (lanes 2–4) as well as the pool of batches 4–6 (lane 5). Fractions 3–11 (corresponding to elution volumes between 102 and 126 mL) from the S300HR column of the [E465A,E468A] sTFR mutant were analyzed SDS-PAGE followed by immunoblotting (lanes 6–14, respectively). Note that homodimeric wild type sTFR typically elutes as a broad peak centered at 165 mL.
