 | Acta Crystallographica Section F Acta Crystallographica Section F STRUCTURAL BIOLOGY COMMUNICATIONS |
journal menu
![[Figure 3]](hv5286fig3mag.jpg)
|
|
Figure 3
rTvTS1 is produced and purified as an active enzyme. (a) SDS–PAGE analysis of rTvTS1 production in P. pastoris. Lane 1, sample from the supernatant of an induced culture. Lane M, Prestained Protein Molecular Weight Marker (Fermentas; labelled in kDa). (b) SDS–PAGE analysis of glycosylated rTvTS1 purified via IMAC (lane 1). Deglycosylated rTvTS1 is shown in lane 2. Lane M, Prestained Protein Molecular Weight Marker (labelled in kDa). (c) Trans-sialidase (left panel) and sialidase (right panel) activity assay performed with 25 pmol glycosylated (black) and deglycosylated (orange) rTvTS1. (d) Analytical gel filtration. The experiment was performed on a Superdex 200 16/60 column. The orange and grey traces indicate the elution profiles for deglycosylated rTvTS1 and the Bio-Rad standards (A, bovine thyroglobulin, molecular mass 670 kDa, Rh = 86.0 Å; B, bovine γ-globulin, molecular mass 158 kDa, Rh = 51.0 Å; C, chicken ovalbumin, molecular mass 44 kDa, Rh = 28.0 Å; D, horse myoglobin, molecular mass 17 kDa, Rh = 19.0 Å; E, vitamin B12, molecular mass 1.35 kDa), respectively. The inset shows an SDS–PAGE analysis of the elution peak. Lane M, Prestained Protein Molecular Weight Marker (labelled in kDa); the lanes marked with the asterisk are fractions from the deglycosylated rTvTS1 elution peak. |
![[Figure 3]](hv5286fig3.jpg)
| STRUCTURAL BIOLOGY COMMUNICATIONS |
ISSN: 2053-230X
