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. 2008 Oct;17(10):1748–1760. doi: 10.1110/ps.036798.108

Figure 5.

Figure 5.

SDS Tricine PAGE showing purification and partial protease digestion of the AscEG and AscEFG complexes. (A) Partial protease digestion of the AscEG complex. (Lane 1) Protein molecular weight marker; (lane 2) His-tagged AscG coexpressed and copurified with AscE to form a soluble AscEG complex. Note that the band for AscG represents AscG (12.93 kDa) with an intact His-tag (1.63 kDa); (lane 3) AscEG after partial protease digestion. The intact AscG fragment contains the N-terminal His-tag and the N-terminal 61 residues of AscG (theoretical MW of 8.24 kDa) with a molecular weight of 8.27 kDa (based on MALDI-TOF MS) as confirmed by N-terminal sequencing. (B) Partial protease digestion of the AscEFG complex. (Lane 1) Protein molecular weight marker; (lane 2) His-tagged AscG coexpressed and copurified with AscE and AscF to form a soluble ternary AscEFG complex; (lane 3) AscEFG after partial protease digestion. The intact AscF fragment has a molecular weight of 4.10 kDa (based on MALDI-TOF MS) and contains the C-terminal 35 residues of AscF (theoretical MW of 4.07 kDa) as confirmed by N-terminal sequencing.