Band 25, Heft 2

Phenylalanine hydroxylase (PAH) is the key enzyme in phenylalanine metabolism, catalyzing its oxidative breakdown to tyrosine. Its function in the committed step of amino acid metabolism requires strict regulation. Thus, several regulatory mechanisms are central for an understanding of PAH at the atomistic level. The enzyme is activated by incubation with phenylalanine and inhibited by tetrahydrobiopterin binding. Furthermore, phosphorylation of Ser-16 in the regulatory domain influences enzyme turnover. All major regulatory processes in PAH are connected to the conformational changes within a protein and its oligomeric assembly. The underlying dynamic processes in the enzyme are tackled by a variety of experimental and computational approaches. We especially emphasize the computational approaches, aiming to unravel the changes in the molecular dynamics of PAH that govern allosteric regulation. State-of-the-art molecular dynamics simulations provide access to the conformational transitions in biological macromolecules at the microsecond time scale and beyond. Thus, in silico strategies are promising for the identification of the complex allosteric mechanisms governing PAH activity in vivo .

Quantum theory of atoms in molecules (QT-AIM) allows detailed insight into the electronic structure of molecules by analysis of the gradient vector field of the electronic density distribution function. First results of a QT-AIM analysis of neutral pterin as well as its anionic and cationic forms in aqueous solution are reported based on density functional theory using B3LYP/6–311+G(2d,p)//6–31G(d) level of theory. Besides reporting QT-AIM results of the atomic partial charges and bond orders of the molecules, their electron density functions, Laplacians and electrostatic potential functions are also shown. The results demonstrate the rather extensive delocalization of both negative and positive extra charges predominantly over the pyrimidine moiety of the pterin ring system and allow a precise quantitation of the effects of addition or elimination of a proton on the bonding structure. Quantum chemical techniques in combination with QT-AIM procedures are thus well-suited to describing the bonding structure as well as the major atomic and bonding features of neutral pterin and its cationic and anionic forms. Moreover, the data demonstrate that the chosen level of theory yields chemically reasonable results while not being computationally too expensive, thus providing a sound basis for further theoretical chemical work on pteridines.

Neonatal sepsis in premature infants is difficult to diagnose, and available markers are still varied. Neopterin is a derivative of pyrazino-pyrimidine produced by macrophages when stimulated by interferon γ. This study aimed to measure and compare the concentration of serum neopterin between groups of neonates with and without sepsis, and its correlation with Töllner’s sepsis score (TSS). This analytic observational study with a cross-sectional design included 46 premature infants with (n=23) and without neonatal sepsis (n=23). Data analysis using the Mann-Whitney U-test was performed to compare the levels of neopterin in premature infants with and without neonatal sepsis, and the Spearman rank correlation test to determine the correlation between neopterin levels and TSS. Neopterin levels in the sepsis group were higher than in the non-sepsis group (116 vs. 41.1 nmol/L; p<0.001). This study also showed a positive correlation between neopterin levels in premature infants with neonatal sepsis and TSS (r=0.776, p<0.001). Neopterin levels were higher in premature infants with neonatal sepsis than in those without it and positively correlated with TSS.

Body building is a sport where ultrastructual damage to muscle fibres aids the development of dense muscle layers. Using a new strong cation exchange (SCX) based chromatography technique to measure neopterin and 7,8-dihydroneopterin, we investigated whether this muscle damage caused increased levels of inflammation. Urine samples were collected over eight consecutive mornings from 10 natural competitive body builders. Samples were analysed using SCX high performance liquid chromatography (HPLC) with urine volume corrected for creatinine and specific gravity (SG). The majority of subjects showed large changes in both neopterin and total neopterin (7,8-dihydroneopterin+neopterin) levels, though the mean data for the group showed no significant change over the week. There was no evidence of the high intensity resistance training causing an accumulation of inflammation as the values for all the body builders returned to close to the starting values after 2 days rest. The SCX analysis had an intra-specific viability of 3.04% and the inter-specific viability was 5.42%. Urine volume correction with SG was found to give the same values as using creatinine. Creatinine and specific gravity are both reliable methods for correcting for urine volume while SCX HPLC provides a new means of measuring urinary neopterin and total neopterin.