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. Author manuscript; available in PMC: 2013 Mar 13.
Published in final edited form as: Biochemistry. 2012 Mar 1;51(10):2113–2121. doi: 10.1021/bi3001038

Figure 2.

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.