Relative positions of Tyr145 and Asp145 with respect to the heme.

<p>Dashed lines correspond to the temporary distances in Å between the heme oxygen and the corresponding hydrogen atoms which undergo time variations during the simulation. Occasionally, Tyr145 forms a hydrogen bond to the heme when internal motion of the protein brings Tyr145 closer to the heme. There is no hydrogen bond formed between Asp145 and heme as they are too far apart from each other.</p

Polymorphisms of <i>CYP51A1</i> from Cholesterol Synthesis: Associations with Birth Weight and Maternal Lipid Levels and Impact on CYP51 Protein Structure

<div><p>We investigated the housekeeping cytochrome P450 <i>CYP51A1</i> encoding lanosterol 14α-demethylase from cholesterol synthesis that was so far not directly linked to human disorders. By direct sequencing of <i>CYP51A1</i> in 188 women with spontaneous preterm delivery and 188 unrelated preterm infants (gestational age <37 weeks) we identified 22 variants where 10 are novel and rare. In infants there were two novel <i>CYP51A1</i> variants where damaging effects of p.Tyr145Asp from the substrate recognition region, but not p.Asn193Asp, were predicted by PolyPhen2 and SIFT. This was confirmed by molecular modeling showing that Tyr145Asp substitution results in changed electrostatic potential of the CYP51 protein surface and lengthened distance to the heme which prevents hydrogen bonding. The CYP51 Tyr145Asp mutation is rare and thus very interesting for further structure/function relationship studies. From the 12 identified known variants rs6465348 was chosen for family based association studies due to its high minor allele frequency. Interestingly, this <i>CYP51A1</i> common variant associates with small for gestational age weight in newborns (p = 0.028) and lower blood total cholesterol and low density lipoprotein cholesterol levels in mothers in 2nd trimester of pregnancy (p = 0.042 and p = 0.046 respectively). Our results indicate a new link between a cholesterol synthesis gene <i>CYP51A1</i> and pregnancy pathologies.</p></div

One-Way ANOVA testing for association between listed parameters and SNP genotypes.

<p>*P-value <0.05.</p

Minor allele frequencies and P-value of the exact binomial test of goodness-of-fit comparing MAF of known <i>CYP51A1</i> variants in neonates or mothers to the general population (1000 Genomes Project). Variants have been identified by sequencing.

<p>*P-value <0.05.</p

Experimental design and workflow.

<p>Panels show <i>CYP51A1</i> analysis workflow by direct sequencing (searching for novel functional variants) and the analysis on population level by genotyping of common<i>CYP51A1</i> variants and family based studies.</p

Novel polymorphisms identified in <i>CYP51A1</i> with sequencing approach and PolyPhen 2 and SIFT predictions.

<p>^a Amino acid residue numbering in RefSeq database NP_000777.1 and in UniProt database Q16850;</p

Cholesterol biosynthesis pathway with genes associated with preterm delivery and low birth weight according to Steffen at all [7] and with input from our study.

<p>Cholesterol biosynthesis pathway with genes associated with preterm delivery and low birth weight according to Steffen at all <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0082554#pone.0082554-Steffen1" target="_blank">[7]</a> and with input from our study.</p

Unveiling the interaction profile of rosmarinic acid and its bioactive substructures with serum albumin

Rosmarinic acid, a phytochemical compound, bears diverse pharmaceutical profile. It is composed by two building blocks: caffeic acid and a salvianic acid unit. The interaction profile, responsible for the delivery of rosmarinic acid and its two substructure components by serum albumin remains unexplored. To unveil this, we established a novel low-cost and efficient method to produce salvianic acid from the parent compound. To probe the interaction profile of rosmarinic acid and its two substructure constituents with the different serum albumin binding sites we utilised fluorescence spectroscopy and competitive saturation transfer difference NMR experiments. These studies were complemented with transfer NOESY NMR experiments. The thermodynamics of the binding profile of rosmarinic acid and its substructures were addressed using isothermal titration calorimetry. In silico docking studies, driven by the experimental data, have been used to deliver further atomic details on the binding mode of rosmarinic acid and its structural components.</p

A combined NMR and molecular dynamics simulation study to determine the conformational properties of rat/mouse 35-55 myelin oligodendrocyte glycoprotein epitope implicated in the induction of experimental autoimmune encephalomyelitis

A combined NMR and molecular dynamics simulation study to determine the conformational properties of rat/mouse 35-55 myelin oligodendrocyte glycoprotein epitope implicated in the induction of experimental autoimmune encephalomyeliti

Comparison of Proposed Putative Active Conformations of Myelin Basic Protein Epitope 87−99 Linear Altered Peptide Ligands by Spectroscopic and Modelling Studies: The Role of Positions 91 and 96 in T-Cell Receptor Activation

This work proposes a structural motif for the inhibition of experimental autoimmune encephalomyelitis (EAE) by the linear altered peptide ligands (APLs) [Ala91,96] MBP87-99 and [Arg91,Ala96] MBP87-99 of myelin basic protein. Molecular dynamics was applied to reveal distinct populations of EAE antagonist [Ala91,96] MBP87-99 in solution, in agreement with NOE data. The combination of the theoretical and experimental results led to the identification of a putative active conformation. This approach is of value as no crystallographic data is available for the APL−receptor complex. TCR contact residue Phe89 has an altered topology in the putative bioactive conformations of both APLs with respect to the native peptide, as found via crystallography; it is no longer prominent and solvent exposed. It is proposed that the antagonistic activity of the APLs is due to their binding to MHC, preventing the binding of self-myelin epitopes, with the absence of an immunologic response as the loss of some interactions with the TCR hinders activation of T-cells