scholarly journals l-myo-inositol 1,4,5,6-tetrakisphosphate is present in both mammalian and avian cells

1988 ◽  
Vol 249 (1) ◽  
pp. 271-282 ◽  
Author(s):  
L Stephens ◽  
P T Hawkins ◽  
N Carter ◽  
S B Chahwala ◽  
A J Morris ◽  
...  
Keyword(s):  
High Performance ◽  
Periodic Acid ◽  
Platelet Activating Factor ◽  
Anion Exchange Chromatography ◽  
Total Radioactivity ◽  
Exchange Chromatography ◽  
Murine Bone Marrow ◽  
Cell Extracts ◽  
Inositol 1,4,5 Trisphosphate ◽  
Inositol Tetrakisphosphate

When myo-[3H]inositol-prelabelled primary-cultured murine bone-marrow-derived macrophages were challenged with platelet-activating factor (PAF; 200 ng/ml), there was a rapid (2.5-fold at 10 s) rise in the intracellular concentration of D-myo-[3H]inositol 1,4,5-trisphosphate, followed by a rise in myo-[3H]inositol tetrakisphosphate. myo-[3H]Inositol tetrakisphosphate fractions were isolated by high-performance anion-exchange chromatography from myo-[3H]inositol-prelabelled chick erythrocytes and primary-cultured macrophages. In both cases [3H]iditol and [3H]inositol were the only significant products (greater than 90% of recovered radioactivity) after oxidation to completion with periodic acid, reduction with NaBH4 and dephosphorylation with alkaline phosphatase. The presence of [3H]inositol after this procedure is consistent with the occurrence of [3H]inositol 1,3,4,5-tetrakisphosphate in the cell extracts, whereas [3H]iditol could only be derived from D- or L-inositol 1,4,5,6-tetrakisphosphate. When [3H]inositol tetrakisphosphate fractions obtained from (A) unstimulated macrophages, (B) macrophages that had been stimulated with PAF for 40s or (C) chick erythrocytes were subjected to the above procedure, radioactivity was recovered in these polyols in the following proportions: A, 60-90% in iditol, with 10-40% in inositol; B, total radioactivity increased by a factor of 9.8, 94% being recovered in inositol and 8% in iditol; C, 70-80% in iditol and 20-30% in inositol. [3H]Iditol derived from myo-[3H]inositol tetrakisphosphate fractions from macrophages and chick erythrocytes was oxidized to sorbose by L-iditol dehydrogenase (L-iditol:NAD+2-oxidoreductase, 1.1.1.14) at the same rate as authentic L-iditol. D-[14C]Iditol, derived from D-myo-inositol 1,4,5-trisphosphate, was not oxidized by L-iditol dehydrogenase. This result indicates that the [3H]iditol was derived from L-myo-inositol inositol 1,4,5,6-tetrakisphosphate. The data are consistent with rapid PAF-sensitive synthesis of D-myo-[3H]inositol 1,3,4,5-tetrakisphosphate in macrophages, and demonstrate that L-myo-inositol 1,4,5,6-tetrakisphosphate is synthesized in both mammalian and avian cells. The levels of L-myo-[3H]inositol 1,4,5,6-tetrakisphosphate in primary-cultured macrophages are not acutely sensitive to PAF.

scholarly journals Enzymatic Synthesis and Characterization of Different Families of Chitooligosaccharides and Their Bioactive Properties

2021 ◽  
Vol 11 (7) ◽  
pp. 3212
Author(s):  
Noa Miguez ◽  
Peter Kidibule ◽  
Paloma Santos-Moriano ◽  
Antonio O. Ballesteros ◽  
Maria Fernandez-Lobato ◽  
...  
Keyword(s):  
High Performance ◽  
Chemical Characterization ◽  
Amperometric Detection ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Enzymatic Production ◽  
Bioactive Properties ◽  
Substrate Properties ◽  
Chitin Hydrolysis

Chitooligosaccharides (COS) are homo- or hetero-oligomers of D-glucosamine (GlcN) and N-acetyl-D-glucosamine (GlcNAc) that can be obtained by chitosan or chitin hydrolysis. Their enzymatic production is preferred over other methodologies (physical, chemical, etc.) due to the mild conditions required, the fewer amounts of waste and its efficiency to control product composition. By properly selecting the enzyme (chitinase, chitosanase or nonspecific enzymes) and the substrate properties (degree of deacetylation, molecular weight, etc.), it is possible to direct the synthesis towards any of the three COS types: fully acetylated (faCOS), partially acetylated (paCOS) and fully deacetylated (fdCOS). In this article, we review the main strategies to steer the COS production towards a specific group. The chemical characterization of COS by advanced techniques, e.g., high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and MALDI-TOF mass spectrometry, is critical for structure–function studies. The scaling of processes to synthesize specific COS mixtures is difficult due to the low solubility of chitin/chitosan, the heterogeneity of the reaction mixtures, and high amounts of salts. Enzyme immobilization can help to minimize such hurdles. The main bioactive properties of COS are herein reviewed. Finally, the anti-inflammatory activity of three COS mixtures was assayed in murine macrophages after stimulation with lipopolysaccharides.

Anion-exchange chromatography of proteins on AG MP-1 using high-performance liquid chromatography equipment

1984 ◽  
Vol 142 (2) ◽  
pp. 373-377 ◽  
Author(s):  
John T. Axelson ◽  
James W. Bodley ◽  
Jeou-Yuan Chen ◽  
Patricia C. Dunlop ◽  
Luann P. Rosenthal ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Anion Exchange ◽  
High Performance ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography
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