Figure 4.

Representative tryptic digestions (A) and SEC profiles (B) of TgCEN1-C and TgCEN2-C in the presence of EGTA or CaCl₂. (A) Tryptic digestion: the intensity of the untreated centrin band in lane 2 was assumed to be 100%. Lane 1: MW marker, lanes 2–9: trypsin digestion products obtained following incubation of protein with trypsin 1:500 w/w for 0, 1, 5, 10, 20, 60, 90 and 120 min, respectively. (B) SEC profiles: proteins were analyzed at 2 mg/mL on a Superdex 75 Increase 10/300 GL gel filtration column in buffer containing EGTA (5 mM; solid trace) or CaCl₂ (5 mM; dotted trace). Inset, calibration curve of distribution coefficient Kd versus the logarithm of the molecular weight. The standard proteins used were: 1, cytochrome c; 2, myoglobin; 3, carbonic anhydrase; 4, ovalbumin; 5, albumin bovine serum.