High-Affinity Fluorogenic Substrate for Tissue Transglutaminase Reveals Enzymatic Hysteresis.

Transglutaminases (TGases) are a family of calcium-dependent enzymes primarily known for their ability to cross-link proteins. Transglutaminase 2 (TG2) is one isozyme in this family whose role is multifaceted. TG2 can act not only as a typical transamidase through its catalytic core but also as a G-protein via its GTP binding site. These two discrete activities are tightly regulated by both environmental stimuli and redox reactions. Ubiquitously expressed in humans, TG2 has been implicated in numerous disease pathologies that require extensive investigation. The catalytic activity of TG2 can be monitored through various mechanisms, including hydrolysis, transamidation, or cleavage of isopeptide bonds. Activity assays are required to monitor the activity of this isozyme not only for studying its transamidation reaction but also for validation of therapeutics designed to abolish this activity. Herein, we present the design, synthesis, and evaluation of a new TG2 activity substrate based on a previously optimized inhibitor scaffold. The substrate APH7 exhibits excellent affinity, selectivity, and reactivity with TG2 (KM = 3.0 µM). Furthermore, its application also allowed the discovery of unique hysteresis at play within the catalytic activity and inhibition reactivity of TG2.

Electrospun nanofibers of cellulose acetate/metal organic framework-third generation PAMAM dendrimer for the removal of methylene blue from aqueous media.

In this research, magnetic metal-organic framework nanofibers were produced by the electrospinning method. The nanocomposite was functionalized by third generation hyperbranched poly(amidoamine) dendrimer (PAMAM) to improve its dye adsorption efficiency from aqueous media. The characteristics of the synthesized magnetic nanocomposite was determined by Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy (EDS) along with elemental mapping analysis and scanning electron microscopy (SEM). Central composite design (CCD) based on response surface methodology (RSM) was performed to optimize the adsorption variables and the values of coefficient of determination (R2) and adjusted R2 were 0.9837 and 0.9490, respectively. The results obtained demonstrated remarkable properties of the synthesized nanofiber as adsorbent for methylene blue from aqueous solutions with the removal efficiency of 95.37% and maximum methylene blue (MB) adsorption capacity of 940.76 mg g-1 under optimized conditions. In addition, it was shown that kinetics and adsorption isotherm of the dye removal process followed Langmuir and pseudo-second-order models, respectively. Thermodynamic study of the dye removal indicated that the process was spontaneous and favorable at higher temperatures. Also, the reusability study shows favorable dye removal efficiency of 80.67% even after 4 cycles. To investigate the performance of the adsorbent for the removal of MB in real samples, a sewage sample from a local hospital was used. The result showed good efficiency of the adsorbent.

The reactivity of lactyl-oxythiamin implies the role of the amino-pyrimidine in thiamin catalyzed decarboxylation.

It has previously been established that the deprotonated amino substituent of the pyrimidine of thiamin diphosphate (ThDP) acts as an internal base to accept the C2H of the thiazolium in ThDP-dependent enzymes. The amino group has also been implicated in assisting the departure of the aldehydic product formed after loss of CO2 from ketoacid substrates. However, the potential role for the pyrimidine amino group in the key decarboxylation step has not been assessed. Oxythiamin contains a hydroxyl group in place of the pyrimidine amino group in thiamin, providing a basis for comparison of reactivity. Lactyl-oxythiamin (LOTh), the conjugate of pyruvic acid and oxythiamin was prepared by condensation of ethyl pyruvate and hydroxyl-protected oxythiamin followed by deprotection and acidic hydrolysis of the ethyl ester. The rate constants observed for the decarboxylation of LOTh in neutral and acidic solutions are about four times smaller than those for the corresponding compound that contains the amino group, lactylthiamin. The difference in reactivity is consistent with the amino group's participation in facilitating the decarboxylation step by allowing a competitive addition pathway that produces bicarbonate and has implications for the corresponding enzymic reaction.