[en] Short communication

[en] Short communication

[en] The transcriptional factor Bach1 forms a heterodimer with small Maf family, and functions as a repressor of the Maf recognition element (MARE) in vivo. To investigate the involvement of Bach1 in the heme-dependent regulation of the expression of the α-globin gene, human erythroleukemia K562 cells were cultured with succinylacetone (SA), a heme biosynthetic inhibitor, and the level of α-globin mRNA was examined. A decrease of α-globin mRNA was observed in SA-treated cells, which was restored by the addition of hemin. The heme-dependent expression of α-globin occurred at the transcriptional level since the expression of human α-globin gene promoter-reporter gene containing hypersensitive site-40 (HS-40) was decreased when K562 cells were cultured with SA. Hemin treatment restored the decrease of the promoter activity by SA. The regulation of the HS-40 activity by heme was dependent on the NF-E2/AP-1 (NA) site, which is similar to MARE. The NA site-binding activity of Bach1 in K562 increased upon SA-treatment, and the increase was diminished by the addition of hemin. The transient expression of Bach1 and mutated Bach1 lacking CP motifs suppressed the HS-40 activity, and cancellation of the repressor activity by hemin was observed when wild-type Bach1 was expressed. The expression of NF-E2 strengthened the restoration of the Bach1-effect by hemin. Interestingly, nuclear localization of Bach1 increased when cells were treated with SA, while hemin induced the nuclear export of Bach1. These results indicated that heme plays an important role in the induction of α-globin gene expression through disrupting the interaction of Bach1 and the NA site in HS-40 enhancer in erythroid cells

[en] Expressions appropriate for the evaluation of the hyperfine splitting in ferric-hemoglobin hydroxide have been derived in the Hartree-Fock approximation using ''giant'' multielectron molecular-orbital wave functions formed as linear combinations of atomic orbitals of the constituent atoms and the molecular orbitals of the OH^- complex incorporating overlap effects. Core orbitals as well as valence orbitals have been considered. The relevant multicenter matrix elements have been evaluated accurately by means of the analytical expressions which we have derived by employing a general closed-form expression developed previously for the coefficients of the expansion of a Slater orbital from one center onto the other. Various calculated electric field gradient components have been analyzed separately in terms of the valence and the core orbitals of the central ion (iron) interacting with the orbitals of the other constituent atoms and the complex OH^-. One finds that the ''local'' and ''distant'' parts of the field gradient due to OH^- nearly cancel one another and, consequently, produce negligible effect on the hyperfine splitting of iron. The nitrogens of the porphyrin plane contribute dominantly to the splitting. Other surrounding atoms contribute less and their influence decreases rapidly as their distance from the central ion increases. On combining various contributions, the calculated hyperfine splitting comes out to be 1.44 + 0.16 mm/sec, which agrees excellently with the experimental splitting 1.57 mm/sec observed by Lang and Marshall. The results from the present calculations have been compared with those obtained by Weissbluth and Maling employing semiempirical treatment on a porphyin-hydroxide model compound. Their estimate is found to give negative sign to the splitting in contrast with our result. Sources of disagreement have been pointed out. Positive hyperfine splitting in ferric-hemoglobin compounds is predicted

[en] Recently, several members of a vertebrate protein family containing a six trans-membrane (6TM) domain and involved in apoptosis and cancer (e.g. STEAP, STAMP1, TSAP6), have been identified in Golgi and cytoplasmic membranes. The exact function of these proteins remains unknown. We related this 6TM domain to distant protein families using intermediate sequences and methods of iterative profile sequence similarity search. Here we show for the first time that this 6TM domain is homolog to the 6TM heme binding domain of both the NADPH oxidase (Nox) family and the YedZ family of bacterial oxidoreductases. This finding gives novel insights about the existence of a previously undetected electron transfer system involved in apoptosis and cancer, and suggests further steps in the experimental characterization of these evolutionarily related families

[en] Studies of irradiation induced lipid peroxidation in fatty acid micelles, both from our own lab and others, are briefly summarized. Steady state measurements have shown the dependence of hydroperoxide yield on the state of aggregation in the lipid and the degree to which the reactive sites are close packed. Chromatographic measurements obeyed the square root dependence of yield on dose rate confirming the proposed chain mechanism. Application to antioxidant studies have demonstrated the highly efficient blockage of the peroxidation chain by α-tocopherol and the subsequent prooxidant effect of the product formed. Time resolved studies have been used to determine rate information for .OH-lipid interaction, radical transfer within the lipid, radical peroxidation, lipid radical movement across the micellar boundary, chain termination, and radical interaction with α-tocopherol. Complimentary laser studies have demonstrated, in contrast to .OH behavior, the comparatively high degree of selectively exhibited by alkoxy radicals toward allylic lipid sites

[en] Crystal structures of MauG in complexes with MADH with its TTQ cofactor in the quinone and quinol forms are reported. MauG has been cocrystallized with methylamine dehydrogenase (MADH) with its TTQ cofactor in the o-quinol (TTQ_OQ) and quinone (TTQ_OX) forms and the structures of the resulting complexes have been solved. The TTQ_OQ structure crystallized in either space group P2₁ or C2, while the TTQ_OX structure crystallized in space group P1. The previously solved structure of MauG in complex with MADH bearing an incompletely formed TTQ cofactor (preMADH) also crystallized in space group P1, although with different unit-cell parameters. Despite the changes in crystal form, the structures are virtually identical, with only very minor changes at the protein–protein interface. The relevance of these structures with respect to the measured changes in affinity between MauG and various forms of MADH is discussed

[en] Biogenic amine-binding proteins mediate the anti-inflammatory and antihemostatic activities of blood-feeding insect saliva. The structure of the amine-binding protein from R. prolixus reveals the interaction of biogenic amine ligands with the protein. Proteins that bind small-molecule mediators of inflammation and hemostasis are essential for blood-feeding by arthropod vectors of infectious disease. In ticks and triatomine insects, the lipocalin protein family is greatly expanded and members have been shown to bind biogenic amines, eicosanoids and ADP. These compounds are potent mediators of platelet activation, inflammation and vascular tone. In this paper, the structure of the amine-binding protein (ABP) from Rhodnius prolixus, a vector of the trypanosome that causes Chagas disease, is described. ABP binds the biogenic amines serotonin and norepinephrine with high affinity. A complex with tryptamine shows the presence of a binding site for a single ligand molecule in the central cavity of the β-barrel structure. The cavity contains significant additional volume, suggesting that this protein may have evolved from the related nitrophorin proteins, which bind a much larger heme ligand in the central cavity

No abstract available

[en] Effects of different levels of soil moisture on growth up to maturity of two cultivars of Pennisetum americanum L. i.e. Tiftlate synthate and IC-8206 were investigated in pots containing sandy clay loam soil. There were four treatments each with six replicates. The survival and appearance of plants was better in response to moderate moisture levels. This shows the sensitivity of the test cultivars to water logging and drought. However, variations were observed in response to these stresses. The cultivars Tiftlate synthate and IC-8206 were found to be truly drought sensitive. Both the cultivars were found to be intermediate in their tolerance to waterlogged conditions. (author)