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Fig. 3
Characterization of cytosolic TMPK. A Partial purification of zebrafish TMPK by DEAE chromatography using cytosolic proteins isolated from zebrafish embryos harvested at 24 hpf and 120 hpf. The bound proteins were eluted with stepwise increase of KCl concentration (from 50 to 250?mM) in DEAE buffer. Fractions were collected and assayed for TMPK activity; B Protein concentration of the corresponding DEAE fractions; C Identification of the canonical TMPK proteins in the TMPK activity peak fractions by western blot analysis using a zebrafish Dtymk-specific antibody. Fractions F16, F18, F20, F21, F25, and F28 from DEAE chromatography were used and only part of the image is shown; D Size exclusion chromatography of partially purified zebrafish TMPK. Fractions F20 and F21 from the DEAE chromatography were used. Fractions were collected and assayed for TMPK activity. The Y-axis on the left side corresponded the TMPK activities from 120 hpf embryos and the Y-axis on the right side corresponded the TMPK activities from 24 hpf embryos. The elution positions of molecular weight markers are indicated