Journal: The Journal of Biological Chemistry

Article Title: Structural basis of EGF-repeat O -glucosylation by the protein O -glucosyltransferase POGLUT2

doi: 10.1016/j.jbc.2026.111361

Figure Lengend Snippet: Functional and structural characterization of human POGLUT2 . A , O -glucosylation sites within epidermal growth factor–like (EGF) repeats. POGLUT1 (Rumi in Drosophila ) adds O -Glc to serine within the C 1 –C 2 consensus sequence C 1 -X- S -X-(P/A)-C 2 . POGLUT2/3 adds O -Glc to serine residue within the C 3 –C 4 consensus sequence C 3 -X-N-T-X-G- S -F-X-C 4 . EOGT (EGF domain–specific O -GlcNAc transferase) catalyzes O -GlcNAc addition to serine or threonine within the C 5 –C 6 consensus sequence C 5 -X-X-G-X-( S/T )-G-X-X-C 6 . Six conserved cysteine residues (C1–C6) form three disulfide bonds, indicated by solid lines . B , sequence alignment of representative EGF-repeat substrates of POGLUT2/3 (human Notch3 EGF10, hN3EGF10; human Notch1 EGF11, hN1EGF11; human Notch4 EGF11, hN4EGF11), POGLUT1 (human Notch1 EGF12 and 13, hN1EGF12 and hN1EGF13; human factor IX EGF, hFA9EGF) and EOGT (human Notch1 EGF20, hN1EGF20). C , enzymatic activity of POGLUT2 with different EGF repeats as acceptor substrates (hN3EGF10, hN1EGF11, and hN4EGF11). hFA9EGF (POGLUT1 substrate) and hN1EGF20 (EOGT substrate) serve as negative control. Data represent mean ± SD from three independent assays. D , preparation of homogeneous POGLUT2. SDS-PAGE analysis shows removal of heterogeneous N -glycans by Endo Hf digestion. The resulting deglycosylated POGLUT2 was further purified by gel-filtration chromatography (Superdex 200 Increase 10/300 GL). E , schematic of the domain organization of POGLUT2. Three pairs of disulfide bonds are depicted by yellow lines . F , overall structure of POGLUT2. Cartoon representation highlights A-domain ( green ), B-domain ( pink ), and filamin domain ( dark gray ). The disulfide bonds are depicted as cyan sticks . G , surface representation of POGLUT2 in the top view . H , close-up view of interdomain interactions between the filamin domain and A-domain, as indicated by the box in ( F ). POGLUT2, protein O-glucosyltransferase 2.

Article Snippet: To remove N -glycans, the target protein was incubated with maltose-binding protein (MBP)–tagged Endo Hf (NEB) at a 1:100 (w/w) enzyme-to-protein ratio overnight at 4 °C.

Techniques: Functional Assay, Sequencing, Residue, Activity Assay, Negative Control, SDS Page, Purification, Filtration, Chromatography

Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–C) PNN neuroprotective role in mitigating EBRO-induced alterations in MBP (A) , NEFL (B) in brain homogenates, and MBP and NEFL levels in CSF (C) in MS rat model. To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–C) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.

Article Snippet: For measuring MBP [E-EL-R0642; Elabscience, Wuhan, China] and NEFL [E-EL-R2536; Elabscience, Wuhan, China] ( ).

Techniques: Standard Deviation, Control