Supplemental Material - An investigation on the use of topical antibiotics for treating eczema and dermatitis in China

Supplemental Material for An investigation on the use of topical antibiotics for treating eczema and dermatitis in China by Juan Shao, Xin Wang, Zhongwen Zhang and Linfeng Li in European Journal of Inflammation.</p

Structural Insights into the Assembly of CARMA1 and BCL10

<div><p>The CBM complex (CARMA1, BCL10 and MALT1) plays a crucial role in B and T lymphocyte activation. CARMA1 serves as a scaffold for BCL10, MALT1 and other effector proteins and regulates various signaling pathways related to the immune response. The assembly of CARMA1 and BCL10 is mediated through a CARD-CARD interaction. Here, we report the crystal structure of the CARD domain of CARMA1 at a resolution of 1.75 Å. The structure consists of six helices, as previously determined for CARD domains. Structural and computational analysis identified the binding interface between CARMA1-CARD and BCL10-CARD, which consists of a basic patch in CARMA1 and an acidic patch in BCL10. Site-directed mutagenesis, co-immunoprecipitation and an NF-κB activation assay confirmed that the interface is necessary for association and downstream signaling. Our studies provide molecular insight into the assembly of CARMA1 and BCL10.</p> </div

The binding surface of CARMA1-CARD.

<p>(<b>A</b>) The representative basic residues (R35, K41, K69 and R72) on the positive surface of the CARMA1-CARD are colored blue. (<b>B</b>) Interactions between the basic residues and surrounding sulfate ions. The side chains of basic residues R35, K41, K69 and R72 and sulfate ions are shown as sticks. The oxygen atoms and sulfur atoms are colored red and yellow, respectively. Hydrogen bonds are shown as red dashed lines. (<b>C</b>) Sequence alignment of CARMA1-CARD proteins from different species. The conserved amino acids are highlighted in red. Conserved residues in the basic patch are denoted with asterisks.</p

Association of CARMA1 and BCL10.

<p>(<b>A</b>) Co-IP analysis of interactions between BCL10-CARD and variants of CARMA1-CARD. HEK293T cells were transiently co-transfected with GFP-tagged BCL10-CARD and wild type or mutants of Myc-tagged CARMA1-CARD constructs. Cell extracts were immunoprecipitated using an anti-Myc antibody and blotted using anti-GFP. (<b>B</b>) Co-IP analysis of interactions between Myc-tagged CARMA1-CARD and wild type or mutants of GFP-tagged BCL10-CARD constructs. (<b>C,D</b>) Bar graph displaying interactions between CARMA1-CARD and BCL10-CARD. (<b>E</b>) The effect of wild type and mutants of CARMA1 on the NF-κB reporter assay. (<b>F</b>) The effect of wild type and mutants of BCL10 on NF-κB activity. RLU: relative luciferase unit; Luc: firefly luciferase activity; and Ren: Renilla luciferase activity. The error bars indicate the standard error of the mean (n = 3 separate experiments). * indicates a P value<0.05, ** indicates a P value<0.001. (<b>G, H</b>) The expression levels of CARMA1 and BCL10 in NF-κB assays were checked by immunoblotting with anti-GFP, anti-MYC and anti-GAPDH antibodys, respectively.</p

Data collection and refinement statistics.

a<p>the highest resolution shell.</p>b<p>.</p>c<p><b><i>R</i></b>_crystal = .</p>d<p><b><i>R</i></b>_free, calculated the same as <b><i>R</i></b>_crystal, but from a test set containing 5% of data excluded from the refinement calculation.</p

The binding surface of BCL10-CARD.

<p>(<b>A</b>) Superposition of homology models of BCL10-CARD. The computational results from the programs MODELLER and SWISS-MODEL are represented with green and blue cartoon models, respectively. (<b>B</b>) The electrostatic surface of BCL10-CARD. Red: negative; blue: positive; and white: neutral. Residues E50, E53 and E54 are labeled. (<b>C</b>) Sequence alignment of BCL10-CARD proteins from different species. The conserved amino acids are highlighted with red, and the conserved acidic residues that make up the acidic patch are denoted with asterisks. (<b>D</b>) Protein docking models of the CARMA1-CARD and BCL10-CARD calculated using the ZDOCK server. CARMA1-CARD and BCL10-CARD are colored magenta and gray, respectively. Three out of ten best scoring complexes place the BCL10-CARD domain approaching the interface containing residues R35, K41, K69 and R72 of CARMA1-CARD. The side chains of residues R35, K41, K69 and R72 are shown as sticks. (<b>E</b>) The interactions between CARD domains in the best docking complex model. CARMA1-CARD and BCL10-CARD are colored magenta and gray, respectively. Side chains of E50, E53 and E54 of BCL10 as well as R53, K41, K69 and R72 of CARMA1 are shown as stick.</p

Formation of Halogenated Polyaromatic Compounds by Laccase Catalyzed Transformation of Halophenols

Laccases are a type of extracellular enzyme produced by fungi, bacteria, and plants. Laccase can catalyze one-electron oxidation of a variety of phenolic compounds using molecular oxygen as the electron acceptor. In this study, transformation of halophenols (XPs) in laccase-catalyzed oxidation processes was explored. We first examined the intrinsic reaction kinetics and found that the transformation of XPs appeared first order to the concentrations of both XPs and laccase. A numerical model was developed to describe the role of humic acid (HA) in this process. It was demonstrated that HA could reverse the oxidation of XPs by acting as the inner filtrator of XP radical intermediates formed upon reactions between the substrates and laccase. The extent of such reversion was proportional to HA concentration. MS analysis in combination with quantum chemistry computation suggested that coupling products were generated. XPs coupled to each via CC or COC pathways, generating hydroxyl polyhalogenated biphenyl ethers (OH-PCDEs) and hydroxyl polyhalogenated biphenyls, respectively. Many of these polyhalogenated products are known to be hazardous to the ecosystem and human health, but they are not synthetic chemicals. This study shed light on their genesis in the environmental media