Conformational integrity of quadruple mutated FLAG-PP6R3. FLAG-tagged PP6R3 wild type (lanes 1-6) and the quadruple mutant E204K, E205K, E259K, E262K (lanes 7-14) were expressed in HEK293T cells, recovered by immunoprecipitation and digested with chymotrypsin for indicated periods of time. Aliquots of the reactions were resolved by SDS-PAGE and immunoblotted for both FLAG (upper panels) and PP6R3 (lower panels). As a control quadruple mutant protein eluted from beads (lane 13) was denatured and digested with the same amount of chymotrypsin for one min (lane 14) and analyzed by immunoblotting. Lines on left edge of panels show migration of molecular size standards.
Guergnon et al. BMC Biochemistry 2009 10:24 doi:10.1186/1471-2091-10-24