A list of OGT substrates identified on kinase substrate peptide microarray.

<p>A list of OGT substrates identified on kinase substrate peptide microarray.</p

A list of OGT substrates identified on nuclear hormone receptor interaction peptide microarray.

<p>A list of OGT substrates identified on nuclear hormone receptor interaction peptide microarray.</p

Identification of human OGT substrates from a kinase substrate and nuclear hormone receptor interaction peptide microarray.

<p>The assay was performed using bacterial lysates containing s-OGT (7 μg/μL), m-OGT (7 μg/μL), or nc-OGT (7 μg/μL), respectively, and in all cases in the presence of UDP-GlcNAc (1 mM). A parallel reaction without UDP-GlcNAc was used as a negative control. Representative images from the kinase substrate peptide microarray (A) and nuclear receptor interaction peptide microarray (B) are shown. Reference spot is highlighted in gray and peptide O-GlcNAcylation by all three isoforms of OGT is highlighted in red. O-GlcNAcylation of each peptide by the three isoforms OGT was quantified and corrected for non-specific signal by subtracting the signal generated without UDP-GlcNAc (from signal with UDP-GlcNAc). Representative heat maps are shown for O-GlcNAcylation of kinase substrate peptides (C) and nuclear receptor interaction peptides (D).</p

The RBL-2_S420A 410–422 peptide inhibited OGT activity.

<p>The inhibitory effect of RBL-2_S420A 410–422 peptide on OGT activity was determined on the nuclear receptor interaction peptide microarray. The reaction was performed by incubating a mixture of purified m-OGT (0.2 μg/μL) and UDP-GlcNAc (0.5 mM) with or without the S420A RBL2 peptide (0.5 mM). The known OGT inhibitor (ST045849) and a no UDP-GlcNAc reaction were used for positive and negative control, respectively. O-GlcNAcylation of NCOA6_1479–1501 peptide (A) and WIPI_1313–318 peptide (B) are shown for the inhibitory effect of RBL-2_S420A 410–422 peptide on OGT activity.</p

Validation of RBL-2_410–422 O-GlcNAcylation.

<p>A, O-GlcNAcylation of RBL-2_410–422 peptide by bacterial lysate containing m-OGT (7 μg/μL) with increasing concentration of UDP-GlcNAc (0–10 mM). B, O-GlcNAcylation of RBL-2_410–422 peptide dependency of increasing total protein concentrations of bacterial lysate containing m-OGT (0, 1.7, 3.5, 7 μg/μL) at 1 mM UDP-GlcNAc. C, O-GlcNAcylation of RBL-2_410–422 peptide by purified m-OGT (0.2 μg/μL) with 1 mM UDP-GlcNAc was inhibited by a known OGT inhibitor (ST045849, 0–200 μM). D, Km value for UDP-GlcNAc was determined with fixed saturating concentration of RBL-2_410–422 peptide, purified m-OGT (0.2 μg/μL) and varying concentration of UDP-GlcNAc (0–2 mM). The Km value derived from the fit to Michaelis-Menten model is 24 μM.</p

Additional file 2 of Variants of BEST1 and CRYBB2 cause a complex ocular phenotype comprising microphthalmia, microcornea, cataract, and vitelliform macular dystrophy: case report

Additional file 2

Coupled Interface and Oxygen-Defect Engineering in Co₃O₄/CoMoO₄ Heterostructures toward Active Oxidation of Ethylbenzene

The catalytic oxidation of ethylbenzene (EB) is a promising route to produce acetophenone (AcPO). Unfortunately, it remains a great challenge to achieve the highly efficient oxidation of EB under solvent-free conditions using molecular oxygen as the sole oxidant. In this contribution, we present a facile strategy to construct hierarchical oxygen vacancy-rich Co3O4/CoMoO4 heterostructures (Vö-CCMO), which delivers a high yield value of 74.5% at 83.2% conversion of EB and selectivity of 89.6% to AcPO. Both experimental studies and theoretical calculations substantiate the important role of oxygen-defect engineering triggered by the modified chemistry environment at the interfaces between the biphasic phases, which contributes to the good catalytic performance. This work illustrates a promising paradigm for the exploit of advanced catalysts toward boosting EB oxidation reaction in a more practical way

S420A is a possible O-GlcNAc site in the RBL-2 protein.

<p>A, peptide mutant used for an Ala scan. For immobilization purposes, peptides were prepared with an extra CG at the N-terminus and Cys 415 was replaced by Ala. B, OGT activity against RBL-2_410–422 peptide mutants was determined using peptide microarray analysis with 0.2 μg/μL purified m-OGT and 1 mM UDP-GlcNAc. C, UDP-Glo assay was used to measure O-GlcNAcylation of RBL-2_410–422 peptide mutants as well. D, kinetic signals from the same microarray experiment of panel B are shown.</p

Additional file 1 of Variants of BEST1 and CRYBB2 cause a complex ocular phenotype comprising microphthalmia, microcornea, cataract, and vitelliform macular dystrophy: case report

Additional file 1

Characteristics of homology search for <i>Athetis dissimilis</i> unigenes.

<p>The number of unigenes matching the top ten species using BlastX in the Nr database is indicated in square brackets</p