Figure 1:

Purification of full-length, human GRK5 expressed in E. coli. A) Map of human GRK5 in a modified pMalC2H10T vector (pEL17). B) Comparison of small-scale expression tests with either Triton X-100 or DNase added to the lysis buffer. GRK5 in the elution is indicated with an arrow. C) Coomassie stain showing the purity after IMAC (1), ion exchange (2) and SEC (3). GRK5 is indicated with an arrow. D) Representative ion exchange chromatogram from a 1 ml HiTrap S column. The column was eluted with a linear gradient from buffer A containing 100 mM NaCl to buffer B containing 500 mM NaCl. GRK5 is eluted with a wide salt concentration ranging from 200 mM NaCl to 500 mM NaCl. E) Representative SEC chromatogram from the indicated region of D) on an analytical S200 column.