Design, synthesis, and activity of analogues of phosphinothricin as inhibitors of glutamine synthetase.

A new group of potent inhibitors of glutamine synthetase was designed and synthesized. The X-ray structure of bacterial glutamine synthetase complexed with phosphinothricin was used for computer-aided structure-based design of the inhibitors, in which the methyl group of phosphinothricin was chosen as the modification site. Amino and hydroxyl moieties were introduced into the phosphinic acid portion of the lead molecule to interact with ammonium binding site in the active cleft of the enzyme. Designed compounds were synthesized in enantiomerically pure form analogous to L-glutamic acid. In vitro kinetic studies with Escherichia coli glutamine synthetase confirmed the biological activity of the designed inhibitors, which with Ki values in the micromolar range (Ki = 0.59 microM for the most potent compound 2) appear to be slightly weaker inhibitors or equipotent to phosphinothricin

Phosphinothricin analogues as inhibitors of plant glutamine synthetases.

A series of phosphinothricin derivatives with a modified methyl group, designed on the basis of the crystal structure of the complex formed by the inhibitor and the target enzyme from Salmonella typhimurium, were evaluated as potential inhibitors of plant glutamine synthetase. These compounds were previously shown to be equipotent or slightly weaker inhibitors to the lead compound against the bacterial enzyme. Because of the presence in higher plants of at least two enzyme forms with different subcellular localization and possible separate metabolic functions, plastidial and cytosolic glutamine synthetases were purified to electrophoretic homogeneity from spinach chloroplasts and cultured tobacco cells, respectively. Kinetic analysis confirmed the ability of the phosphinothricin analogues to inhibit both isoenzymes in the micromolar range, with a mechanism of a competitive type with respect to glutamate. Interestingly, some of them exerted a differential effect against either the two plant isoforms, or against the plant versus the bacterial enzyme

Isolation and characterization of two new microbial strains capable of degradation of the naturally occurring organophosphonate–ciliatine

Air-born mixed fungal and bacterial culture capable of complete degradation of ciliatine was isolated. The utilization of the natural organophosphonate proceeded in the phosphate independent manner. Enzymatic activity involved in ciliatine degradation studied in the fungal cellfree extract proved to be distinct from bacterial pathway described before

Herbicidal pyridyl derivatives of aminomethylene-bisphosphonic acid inhibit plant glutamine synthetase.

A series of aminomethylene-bisphosphonic acid derivatives, previously synthesized and shown to be endowed with herbicidal properties, were evaluated as potential inhibitors of plant glutamine synthetase. The cytosolic form of the enzyme was partially purified from rice cultured cells and assayed in the presence of millimolar concentrations of the compounds by means of three different assay methods, respectively measuring the hemibiosynthetic, the transferase, and the full biosynthetic reactions. Several compounds were found to exert a remarkable inhibition, with I50 values similar to those obtained under the same conditions with a well-established inhibitor of glutamine synthetase, the herbicide phosphinothricin. Contrary to the reference compound, enzyme kinetics accounted for a reversible inhibition mechanism. The biological activity of the most active derivatives was further characterized by measuring free glutamine levels in cell suspension rice cultures following treatment with the inhibitors. Results confirmed their ability to interfere in vivo with nitrogen metabolism. A preliminary analysis of structure-activity relationship allowed it to be hypothesized that steric rather than electronic factors are responsible for the inhibitory potential of these compounds