Fig.5. Effects of temperature on the ATPase activity of unstimulated and RCMLA-stimulated Hsc70 purified from Gillichthys mirabilis white muscle. Assays, containing 2µg of Hsc70 plus 3.7x10⁵Bq of [-³²P]ATP in assay buffer, were incubated in the absence (open circles) or presence (triangles) of 80µmoll^–1 RCMLA at 5°C increments in a temperature gradient that ranged from 10 to 45°C. Following a 60min incubation, triplicate 1µl samples were removed from each assay tube and analyzed for ATP hydrolysis by thin-layer chromatography. The black circles represent a control assay containing only 80µmoll^-1 RCMLA. Each data point represents the mean pixel absorbance volume (background-corrected) ± S.D. (N=3). See Materials and methods for a full description of the assay buffer components.