Synthesis of 9-β-d-Glucopyranosyl-Adenine-6′-Phosphate

Synthesis of 9-β-d-glucopyranosyl-adenine-6′-phosphate is described. The method developed here involves the process of condensation of base (chloromercuri-6-benzamidopurine) (I) with phosphorylated sugar (2,3,4-tri-O-acetyl-6-diphenylphosphoryl-α-d-glucopyranosyl bromide) (II). This reaction gives crystalline 6-benzamido-9-(2′,3′,4′-tri-O-acetyl-6′-diphenylphosphoryl-β-d-glucopyranosyl)-purine (III) in high yield, which is converted to the desired nucleotide by alkaline hydrolysis.     

Synthesis and Juvenile Hormone Activity of Some Terpenoid Phenyl Ethers

Thirty eight new compounds with juvenile hormone (JH) activity were synthesized and evaluated biologically on Bombyx mori L. Among them, the activities of 7,8-epoxy-4,8-dimethyl-1 -(p-ethylphenoxy)-3-undecene and 7,8-epoxy-4,8-dimethyl-l-(p-methylphenoxy)-3-undencene were proved to be more than two thousand times as high as that of the C₁₈-Cecropia JH (mixed isomers). Structure-activity relationship of these compounds was discussed.     

Synthesis of Isocoumarins

A synthesis of oospolactone (3,4-dimethyl-8-hydroxyisocoumarin), a metabolite of Oospora astringenes, is described.     

Synthesis and Biological Activity of Analogs of the Cecropia Juvenile Hormone with Different Alkyl Substituents at C-7 and C-11

Sixteen new Cecropia juvenile hormone (JH) analogs with different alkyl substituents at C–7 and C–11 were synthesized as stereoisomeric mixtures. The epoxides with n-propyl or n-butyl and methyl groups at C-11 and methyl or ethyl group at C-7 showed high JH activity on Bombyx mori L. Structure-activity relationship of the JH analogs was discussed.     

Phosphorylated TandeMBP: A unique protein substrate for protein phosphatase assay

To analyze a variety of protein phosphatases, we developed phosphorylated TandeMBP (P-TandeMBP), in which two different mouse myelin basic protein isoforms were fused in tandem, as a protein phosphatase substrate. P-TandeMBP was prepared efficiently in four steps: (1) phosphorylation of TandeMBP by a protein kinase mixture (Ca²⁺/calmodulin-dependent protein kinase Iδ, casein kinase 1δ, and extracellular signal-regulated kinase 2); (2) precipitation of both P-TandeMBP and protein kinases to remove ATP, Pi, and ADP; (3) acid extraction of P-TandeMBP with HCl to remove protein kinases; and (4) neutralization of the solution that contains P-TandeMBP with Tris. In combination with the malachite green assay, P-TandeMBP can be used to detect protein phosphatase activity without using radioactive materials. Moreover, P-TandeMBP served as an efficient substrate for PPM family phosphatases (PPM1A, PPM1B, PPM1D, PPM1F, PPM1G, PPM1H, PPM1K, and PPM1M) and PPP family phosphatase PP5. Various phosphatase activities were also detected with high sensitivity in gel filtration fractions from mouse brain using P-TandeMBP. These results indicate that P-TandeMBP might be a powerful tool for the detection of protein phosphatase activities.     

A high molecular weight glutamyl endopeptidase and its endogenous inhibitors from cucumber leaves

We purified a glutamyl endopeptidase that is a major foliar endopeptidase in cucumber. The endopeptidase had a molecular mass of 400 kDa, consisted of four subunits of 97 kDa, and was inactivated by SH-modifying reagents. Its optimum pH and optimum temperature were 8.0 and 30-37 degrees C, respectively. An internal amino acid sequence of the endopeptidase was highly homologous to a partial sequence of unidentified proteins deduced from genetic information for Arabidopsis thaliana, soybean and rice, but not to the sequences of bacterial glutamyl endopeptidases or animal proteases. Therefore, the unidentified proteins might be glutamyl endopeptidases and be widely distributed only among plant species. The activity of the cucumber glutamyl endopeptidase was inhibited by at least three inhibitors existing in cucumber leaves. One of the inhibitors was a competitive inhibitor of 25 kDa, which did not significantly inhibit commercial endopeptidases derived from animals and microorganisms. This suggests that the cucumber glutamyl endopeptidase might be controlled by endogenous inhibitors in vivo.