Purification and properties of cyclic AMP-dependent protein kinase from rat epididymis

Two protein kinases (I and II: EC 2.7.1.37) that show a high degree of substrate specificity for protamine rather than histones, phosvitin and casein were partly purified from rat epididymal tissue. The enzymes were present in the cytosol because greater than 80% of the enzymic activity was recovered in the soluble fraction. The kinases required Mg2+ for activity although Co2+ and Mn2+ were partial substitutes. Zn2+ (1 mM) inhibited nearly completely the activity of the enzymes. Both the kinases showed high affinity for activation with cyclic AMP compared to other cyclic nucleotides. Amino acid analysis of 32P-labelled protamine product revealed that the kinases transfer the terminal phosphate of ATP to serine residues of the protein. The isoenzymes I and II showed certain differences in relation to their hydroxyapatite-chromatography profiles, pH activation profiles, heat sensitivity and Km for ATP and cyclic AMP.     

Two forms of phosphoinositol kinase from germinating mung bean seeds

Phosphoinositol kinase, the key enzyme responsible for the biosynthesis of higher inositol phosphates has been isolated from the cotyledons of mung beans germinated for 24 hr and has been resolved into two different forms, phosphoinositol kinase A and phosphoinositol kinase B. Both forms were purified to homogeneity and characterized. The K_m values for ATP with phosphoinositol kinase A (1.78 × 10^?4 M) and phosphoinositol kinase B (3.12 × 10 ^?5 M) showed that phosphoinositol kinase B had a greater affinity for ATP. ATP could be partially replaced as phosphate donor by UTP and phosphoenolpyruvate in the case of phosphoinositol kinase A but not in the case of phosphoinositol kinase B.     

Xanthone and flavonol constituents of Swertia hookeri

The whole plant of Swertia hookeri, collected at flowering has been shown to contain two tri- and nine tetraoxygenated free, glucosyloxy, and stearyl ester xanthones and one flavonol stearyl ester. Among these, three are previously unreported in nature and one was known previously only as a synthetic compound. The xanthones are based on 1,3,5,-, 1,3,5,8- and 1,3,7,8-oxygenated systems with the middle oxygenation pattern predominating. The two ester compounds appeared only at the flowering stage. Plants collected at the pre-flowering stage gave the corresponding free compounds. The biochemical and biological significance of these findings are appraised.     

Induction of N-acetylmannosamine catabolic pathway in yeast.

N-Acetylmannosamine kinase activity is absent from yeast cells grown on N-acetylmannosamine. However, other enzymes of the catabolic pathway, namely, N-acetylmannosamine-2-epimerase, N-acetylglucosamine kinase and glucosamine-6-phosphate deaminase are induced. In addition, a high affinity uptake system (permease) for the uptake of N-acetylglucosamine is synthesized under these conditions. The presence of either N-acetylmannosamine or N-acetylglucosamine as inducer is essential for the induced synthesis of these enzymes. The enzyme synthesis stops and their concentration in the cells declines rapidly as soon as inducer is removed from the medium. N-Acetyl-D-galactosamine can also induce all these enzymes except for N-acetylmannosamine-2-epimerase, suggesting the convergence of catabolic pathways for both the aminosugars at N-acetyl-D-glycosamine. Experiments with inhibitors of macromolecule synthesis suggest that the snythesis of RNA and protein is necessary for the induction of these cyzymes whereas the synthesis of DNA is not.     

RNA synthesis in maturing avian erythrocyte nuclei

The rate of RNA synthesis and its inhibition by α-amanitin in the nuclei of mature and immature avian erythrocytes are increased with the increase in ionic strength of incubation medium. Polyacrylamide gel electrophoresis indicates that heterogeneous species of RNAs are synthesised in the mature and immature erythrocyte nuclei. However, a large number of high molecular weight RNAs are synthesised in the nuclei of immature erythrocytes. Elution profiles on poly(U)-sepharose chromatography indicate that the RNAs synthesised in the nuclei of two types of cell contain poly(A) segments. Sixteen per cent of mature erythrocyte nuclear RNA syntbesised are polyadenylated, while it is 13% in immature erythrocyte nuclei. However, the total RNA synthesised is 2–3 fold higher in immature erythrocyte nuclei than that in mature erythrocyte nuclei.     

Purification and characterization of myo-inositol hexaphosphate-adenosine diphosphate phosphotransferase from Phaseolus aureus

myo-Inositol hexaphosphate adenosine diphosphate phosphotransferase transfers phosphate from myo-inositol hexaphosphate to adenosine diphosphate to synthesize adenosine triphosphate. This enzyme has been isolated and purified from ungerminated mungbean seeds and found to be different from guanosine diphosphate phosphotransferase. A purification of about 200-fold with 15% recovery has been obtained. The optimal pH of the reaction is 7.0 and is dependent on the presence of a divalent cation, i.e., Mg²⁺ and Mn²⁺. The K_m value for myo-inositol hexaphosphate has been found to be 0.41 × 10^?4m and V is 90.0 nmol of P_i transferred per milligram of protein per 20 min. K_m for ADP is 0.88 × 10^-4m and V is 83.3 nmol of phosphorus transferred to ADP per milligram of protein per 20 min. The ADP phosphotransferase reaction is reversible to the extent of about 50% of the forward reaction. dADP is partly effective as an acceptor but other ribonucleoside mono- and diphosphates cannot substitute for ADP. The products ATP and myo-inositol pentaphosphate have been confirmed by several criteria. It has also been shown that this enzyme transfers phosphate only from a specific phosphoryl group (C-2 position) of myo-inositol hexaphosphate for the synthesis of ATP and 1,3,4,5,6-myo-inositol pentaphosphate or pentakis (dihydrogen phosphate).     

Effect of continuous light and darkness on the testicular histology of toad (Bufo melanostictus)

Continuous darkness decreases spermatogenesis as well as Leydig cell function whereas continuous illumination suppresses spermatogenesis along with increased Leydig cell activity.     

Conformational studies on the ABH and Lewis blood group oligosaccharides

The possible conformations for the ABH and Lewis blood group oligosaccharides have been studied by an energy-minimisation procedure using empirical potential functions. It has been found that the conformation of the core structure is not altered significantly by the addition of l-fucose, galactose or N-acetyl galactosamine residues at the non-reducing end. Correlation of the preferred conformations with their known binding properties suggests that the differences between type 1 and type 2 structures become significant only when a large enough fragment of the determinant is considered. It is suggested that non-specific reagents may have small binding sites while the reagents that are specific for type 1 or type 2 structures may have larger binding sites. A two-pocket model has been proposed for antibodies and lectins which can distinguish the A1 and A2 antigens.