ABSTRACT
Proliferative response of human peripheral blood mononuclear cells (PBMC) stimulated by new lectins purified from seeds of differents Artocarpus species from Vietnam (A. asperulus, A. heterophyllus, A. masticata, A. melinoxylus, A. parva and A. petelotii) was studied and compared to those of the lectin jacalin purified from jackfruit (A. heterophyllus) seeds collected in the island La Réunion. All lectins stimulated human PBMC to proliferate, with a variable efficiency of the mitogenic activity. Phenotypic analysis of cells recovered after 7 day-cultures showed that these lectins mostly stimulated CD4+ T lymphocytes. These results suggest that these lectins from different Artocarpus species are similar in terms of their mitogenic activity although their structural features are not identical.
Subject(s)
Fruit/chemistry , Lectins/pharmacology , Mitogens/pharmacology , CD4-Positive T-Lymphocytes/cytology , Cell Division/drug effects , Flow Cytometry , Humans , Lectins/isolation & purification , Leukocytes, Mononuclear/cytology , Plant Lectins , Reunion , VietnamABSTRACT
Jacalins, from jack-fruit seeds of 2 wild species (Artocarpus asperulus, Artocarpus masticata) were purified by mucine-sepharose 4B affinity chromatography. The alpha and beta chains were separated by reverse phase high pressure liquid chromatography (HPLC). Analysis by HPLC with a C8 column and the determination of the N-terminal sequence of the alpha-chain of these jacalins allowed the identification of a new alpha-chain. Immunological cross-reactivity and carbohydrate specificity indicate that jacalins possessing the new alpha-chain conserve structural and functional properties of the other members of Artocarpus genus.
Subject(s)
Interferon Inducers/isolation & purification , Lectins/isolation & purification , Plant Lectins , Amino Acid Sequence , Chromatography, High Pressure Liquid , Interferon Inducers/chemistry , Lectins/chemistry , Peptide Chain Termination, TranslationalABSTRACT
The jacalins of three Artocarpus species were purified by affinity chromatography on a desialylated mucin-CNBr-Sepharose 4B column. The beta-chains and the 14 kDa alpha-chains were separated by high pressure liquid chromatography and the 17 kDa chains by preparative electrophoresis. The 17 kDa and 14 kDa chains had a similar highly conserved N-terminal sequence. The beta-chains were different for the three species and Artocarpus champeden contained two different beta-chains. CNBr cleavage of the 17 kDa polypeptide of Artocarpus tonkinensis yielded one peptide more than the 14 kDa. The N-terminal sequence of this fragment was similar to that of the beta-chain proving that this chain results from a proteolytic cleavage at the C-terminus of the 17 kDa peptide. The large heterogeneity of the beta-chains of jacalins from different species could be used as a marker for evolutionary studies on the Artocarpus family.
Subject(s)
Lectins/chemistry , Plant Lectins , Plant Proteins/chemistry , Protein Precursors/chemistry , Amino Acid Sequence , Amino Acids/analysis , Lectins/isolation & purification , Molecular Sequence Data , Peptides/chemistry , Plant Proteins/isolation & purification , Protein Precursors/isolation & purificationABSTRACT
Various analogs of adenosine 5'-triphosphate with a modified terminal phosphate group have been tested in energy-requiring reactions with intact mitochondria and submitochondrial particles. It is shown that the fluorophosphate analog ATP(gamma F) is a strong inhibitor of mitochondrial respiration and of energy requiring reactions which involve the participation of high energy intermediates, generated aerobically by the respiratory chain. On the other hand, ATP(gamma F) does not affect the ATPase activity of intact or disrupted mitochondria and is less effective in inhibiting ATP-driven reactions. The imidophosphate analog AMP-P(NH)P also inhibits the partial reactions of oxidative phosphorylation, but does not affect ATP synthesis from ADP and Pi. In contrast to ATP(gamma F), it is strong inhibitor of both soluble and membrane-bound mitochondrial ATPases. The biological implication of the complementary effects of ATP(gamma F) and AMP-P(NH)P on mitochondria-catalysed reactions is discussed while suggesting the use of such nucleotide analogs as specific tools for the study of ATP-forming and ATP-utilizing reactions in mitochondria.
Subject(s)
Adenosine Triphosphatases/metabolism , Adenosine Triphosphate/analogs & derivatives , Mitochondria/metabolism , Oxidative Phosphorylation/drug effects , Adenosine Triphosphate/pharmacology , Animals , Brain/metabolism , Dinitrophenols/pharmacology , Kinetics , Mitochondria/drug effects , Mitochondria, Liver/drug effects , Mitochondria, Liver/metabolism , Rats , Structure-Activity RelationshipABSTRACT
A total of 26 different purine nucleotides with specific modifications in the base moiety and/or in the polyphosphate chain as well as various combinations of nucleotides were tested as allosteric effectors of beef liver glutamate dehydrogenase (L-glutamate : NAD(P)+ oxidoreductase (deaminating), EC 1.4.1.3). The capacity of these nucleotide analogs to activate or to inhibit the glutamate dehydrogenase activity is expressed quantitatively and scaled between the extreme effects of ADP and GTP, respectively. The significance of distinct structural elements for the enzyme-effector interaction is discussed. While the inhibitory GTP site is less specific, accepting many natural and most modified nucleoside triphosphates as inhibitors, the activating ADP site shows a much higher specificity for nucleotides as activators.
