Novel Antimicrobial Peptide from the Hepatopancreas of the Red King Crab.

Crustaceans have successfully adapted to survive in their natural habitat, rich in microorganisms, due to the presence of antimicrobial peptides (AMPs) in their organism. They achieve this adaptation despite lacking the highly specific adaptive immune system found in vertebrates. One valuable source of AMPs is the hepatopancreas, a waste product from crab fishery and its processing.

A hydromorphic reevaluation of the forgotten river civilizations of Central Asia.

The Aral Sea basin in Central Asia and its major rivers, the Amu Darya and Syr Darya, were the center of advanced river civilizations, and a principal hub of the Silk Roads over a period of more than 2,000 y. The region's decline has been traditionally attributed to the devastating Mongol invasion of the early-13th century CE. However, the role of changing hydroclimatic conditions on the development of these culturally influential potamic societies has not been the subject of modern geoarchaeological investigations.

Comparative analysis of the active sites of orthologous endolysins of the Escherichia lytic bacteriophages T5, RB43, and RB49.

The methods of solution NMR, circular dichroism (CD), and differential scanning calorimetry (DSC) were used to study two zinc-containing L-alanyl-D-glutamate peptidases - endolysins of the pseudo T-even myoviruses RB43 and RB49 (EndoRB43 and EndoRB49, respectively), which are orthologous to the EndoT5, which is a zinc-containing L-alanyl-D-glutamate peptidase of the T5 siphovirus. The spatial conservation of the Zn-binding sites for the enzymes EndoT5, EndoRB43, and EndoRB49 was established, and the key role of Zn ions in the stabilization of the spatial structures of these three peptidases was confirmed. We are showing here that the binding of the Zn ion in the active center of EndoRB49 peptidase causes conformational rearrangements similar to those observed in the EndoT5 peptidase upon binding of Zn and Ca ions and lead to the formation of a catalytically active form of the enzyme.

On the roles of calcium and zinc ions in the formation of a catalytically active form of the metalloenzyme, l-alanyl-d-glutamate peptidase of the bacteriophage T5 (EndoT5).

Structural consequences of the binding of metal ions (regulatory Ca and catalytic Zn) to the metalloenzyme l-alanyl-d-glutamate peptidase of the bacteriophage T5 (Endo T5) and some of its analogues containing single amino acid substitutions in the active center were analyzed by nuclear magnetic resonance (NMR), circular dichroism (CD) and calorimetry. Analyses revealed that the native EndoT5 undergoes strong structural rearrangements as a result of Zn binding. This structural rearrangement resulting in the formation of an active enzyme is completed by the Ca binding.

Effect of C-terminal His-tag and purification routine on the activity and structure of the metalloenzyme, l-alanyl-d-glutamate peptidase of the bacteriophage T5.

In this work, we studied the effect of the C-terminally attached poly-histidine tag (His-tag), as well as the peculiarities of the protein purification procedure by the immobilized metal affinity chromatography (IMAC) on the activity and structure of the metalloenzyme, l-alanyl-d-glutamate peptidase of bacteriophage T5 (EndoT5), whose zinc binding site and catalytic aspartate are located near the C-terminus. By itself, His-tag did not have a significant effect on either activity or folding of the polypeptide chain, nor on the binding of zinc and calcium ions to the protein. However, the His-tagged EndoT5 samples had low shelf-life, with storage of these samples resulting in an increased propensity for protein self-association and decreased enzymatic activity of EndoT5.