cyclic-gmp and thiophosphoric-acid

An integrated set of reactions and conditions that allow an eight-step one-flask synthesis of the protected derivatives of c-di-GMP and the [R(p),R(p)] and [R(p),S(p)] thiophosphate analogues is reported. Deprotection is also carried out as a one-flask procedure, with the final products isolated by crystallization from the reaction mixture. Chromatography is only used for separation of the thiophosphate diastereomers.

Topics: Cyclic GMP; Kinetics; Phosphates; Stereoisomerism

Seven phosphorothioate analogs of c-di-GMP (all diastereomers of mono-, di-, and trithiophosphates) were prepared to assess the impact of the thioate substitutions on c-di-GMP polymorphism using 1D (1)H and (31)P NMR, along with 2D NOESY and DOSY, for both the Na(+) and K(+) salts. The K(+) salts display more extensive higher order complex formation than the Na(+) salts, resulting primarily in octamolecular complexes with K(+), but tetramolecular complexes with Na(+). Further, the presence of one or two [S(P)] sulfurs specifically stabilizes anti complexes and/or destabilizes syn complexes, while the presence of two [S(P)] sulfurs promotes extensive aggregation.

Topics: Cyclic GMP; Isomerism; Magnetic Resonance Spectroscopy; Molecular Conformation; Phosphates; Potassium; Salts; Sodium

The synthesis of mono- and bisphosphorothioate analogues of 3',5'-cyclic diguanylic acid (cdiGMP) via the modified H-phosphonate chemistry is reported. The immunostimulatory properties of these analogues were compared with those of cdiGMP.

Topics: Bronchi; Chemistry, Pharmaceutical; Chromatography, High Pressure Liquid; Cyclic GMP; Dose-Response Relationship, Drug; Drug Design; Humans; Immune System; Inflammation; Models, Chemical; Neutrophils; Organophosphonates; Phosphates; Vaccines

Thiophosphorylation and phosphorylation of 5% perchloric acid extractable proteins from calf thymus chromatin were studied using a cyclic GMP-dependent protein kinase from bovine lung and a nuclear protein kinase II from rat liver. The phosphorylation reaction catalyzed by nuclear protein kinase II utilized [gamma -35S]ATP as a phosphate donor almost as efficiently as [gamma -32P]ATP, but the cGMP-dependent protein kinase mediated phosphorylation by [35S]ATP was about 20 times less effective than that by [32P]ATP. In addition, using [35S]ATP instead of [32P]ATP changed markedly the cGMP-dependent phosphorylation pattern of the PCA-extractable proteins as examined by gel electrophoresis. Thus, depending on the type of protein kinase, the results from thiophosphorylation and phosphorylation reactions may vary considerably.

Topics: Amino Acids; Animals; Cattle; Cyclic GMP; Electrophoresis, Polyacrylamide Gel; Female; High Mobility Group Proteins; In Vitro Techniques; Male; Phosphates; Phosphorylation; Protein Kinases; Rats; Rats, Inbred Strains; Thymus Gland