The Catalytic Mechanism of Pdx2 Glutaminase Driven by a Cys-His-Glu Triad: A Computational Study

The catalytic mechanism of Pdx2 was studied with atomic detail employing the computational ONIOM hybrid QM/MM methodology. Pdx2 employs a Cys-His-Glu catalytic triad to deaminate glutamine to glutamate and ammonia – the source of the nitrogen of pyridoxal 5’-phosphate (PLP). This enzyme is, therefore, a rate-limiting step in the PLP biosynthetic pathway of Malaria and Tuberculosis pathogens that rely on this mechanism to obtain PLP. For this reason, Pdx2 is considered a novel and promising drug target to treat these diseases. The results obtained show that the catalytic mechanism of Pdx2 occurs in six steps that can be divided into four stages: (i) activation of Cys₈₇, (ii) deamination of glutamine with the formation of the glutamyl-thioester intermediate, (iii) hydrolysis of the formed intermediate, and (iv) enzymatic turnover. The kinetic data available in the literature (19.1–19.5 kcal mol⁻¹) agree very well with the calculated free energy barrier of the hydrolytic step (18.2 kcal.mol⁻¹¹), which is the rate-limiting step of the catalytic process when substrate is readily available in the active site. This catalytic mechanism differs from other known amidases in three main points: i) it requires the activation of the nucleophile Cys₈₇ to a thiolate; ii) the hydrolysis occurs in a single step and therefore does not require the formation of a second tetrahedral reaction intermediate, as it is proposed, and iii) Glu₁₉₈ does not have a direct role in the catalytic process. Together, these results can be used for the synthesis of new transition state analogue inhibitors capable of inhibiting Pdx2 and impair diseases like Malaria and Tuberculosis.     

MPF6D: masked pyramid fusion 6D pose estimation

Pattern Analysis and Applications - Object pose estimation has multiple important applications, such as robotic grasping and augmented reality. We present a new method to estimate the 6D pose of...     

Effect of chitosan-based solutions applied as edible coatings and water glazing on frozen salmon preservation – A pilot-scale study

The aim of this research was to compare the effect of chitosan solutions on frozen salmon preservation with that of water glazing. For this purpose, three chitosan solutions (0.25%, 0.50% and 0.75% w/v) and water were applied in different amounts (6%, 8% and 11% of coated fillet weight) directly on the surface of frozen salmon. In order to accelerate the deterioration processes, salmon was stored during 14 weeks at −5 °C. Microbial and chemical indices were used to assess deterioration during storage and the coating stability was evaluated through weight loss measurements. The results obtained showed that chitosan coatings can be a good barrier to protect frozen fish from deterioration. Microbial growth, assessed by total viable counts (TVC), and total volatile basic nitrogen (TVB-N) were maintained below the maximum limits recommended which are 5 × 105 CFU/g and 35 mg nitrogen/100 g fish, respectively. The use of 0.50% and 0.75% chitosan solutions generally demonstrated to be more efficient in preventing salmon weight loss.