• 분석과학
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• 제7권4호
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• pp.505-515
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• 1994

DMSO(dimethylsuloxide-$d_6$) 용액 속에 존재하는 CN/CN 리간드의 hemin 착물과 CN/DMSO의 hemin 착물이 $^1H$ NMR로 기록되어지고 분석되어졌다. Hemin으로의 CN 착물화 과정은 온도에 따라 변화함을 NMR 스펙트럼이 보여 주며, 한 개의 CN 리간드에서 두 개의 CN 리간드착물로 바뀌는 과정의 열역학함수는 ${\Delta}H^{\circ}=736.6cal/mol$ 및 ${\Delta}S^{\circ}=16.4eu$인 흡열과정을 나타낸다. CN/DMSO의 hemin 착물은 Curie behavior로부터의 벗어남은 high-spin 성격의 존재를 나태내고, 이는 Fe-DMSO 결합이 순간적으로 깨짐을 의미하며, 이러한 CN/DMSO hemin 착물이 한 개의 axial ligand가 약한 heme 단백질의 전자 및 분자구조의 model complex로 작용할 수 있음을 보여 준다.

• 한국미생물·생명공학회지
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• 제49권1호
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• pp.57-64
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• 2021

Lactic acid bacteria (LAB), belonging to the Firmicutes phylum, lack heme biosynthesis and, thus, are characterized as fermentative and catalase-negative organisms. To verify the hypothesis that heme-rich-nutrients might compensate the heme-biosynthesis incapability of non-respiratory LAB in animal gut, a heme-rich-nutrient was fed to a dog and its fecal microbiome was analyzed. Firmicutes abundance in the feces from the heme-rich-nutrient-fed dog was 99%, compared to 92% in the control dog. To clarify the reason of increased Firmicutes abundance in the feces from the heme-rich-nutrient-fed dog, Lacobacillus gasseri were used as model anerobic LAB to study a purified heme (hemin). The anaerobic growth of L. gasseri in the medium with 25 µM hemin supplementation was faster than that in the medium without hemin, while the growth in the 50 µM hemin-supplemented medium did not vary. Cellular activities of the cytochrome bd complex were 1.55 ± 0.19, 2.11 ± 0.14, and 2.20 ± 0.08 U/gcell in the cells from 0, 25, and 50 µM hemin-supplemented medium, while intracellular ATP concentrations were 7.90 ± 1.12, 11.95 ± 0.68, and 12.56 ± 0.58 µmol_ATP/g_cell, respectively. The ROS-scavenging activities of the L. gasseri cytosol from 25 µM and 50 µM hemin-supplemented medium were 68% and 82% greater than those of the cytosol from no hemin supplemented-medium, respectively. These findings indicate that external hemin could compensate the heme-biosynthesis incapability of L. gasseri by increasing the cytosolic ROS-scavenging and extra ATP generation, possibly through increasing the electron transfer. Increase in the number of anaerobic bacteria in heme-rich-nutrient-fed animal gut is discussed based on the results.

• Molecules and Cells
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• 제45권8호
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• pp.575-587
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• 2022

Human ABCB6 is an ATP-binding cassette transporter that regulates heme biosynthesis by translocating various porphyrins from the cytoplasm into the mitochondria. Here we report the cryo-electron microscopy (cryo-EM) structures of human ABCB6 with its substrates, coproporphyrin III (CPIII) and hemin, at 3.5 and 3.7 Å resolution, respectively. Metal-free porphyrin CPIII binds to ABCB6 within the central cavity, where its propionic acids form hydrogen bonds with the highly conserved Y550. The resulting structure has an overall fold similar to the inward-facing apo structure, but the two nucleotide-binding domains (NBDs) are slightly closer to each other. In contrast, when ABCB6 binds a metal-centered porphyrin hemin in complex with two glutathione molecules (1 hemin: 2 glutathione), the two NBDs end up much closer together, aligning them to bind and hydrolyze ATP more efficiently. In our structures, a glycine-rich and highly flexible "bulge" loop on TM helix 7 undergoes significant conformational changes associated with substrate binding. Our findings suggest that ABCB6 utilizes at least two distinct mechanisms to fine-tune substrate specificity and transport efficiency.

• Asian-Australasian Journal of Animal Sciences
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• 제18권1호
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• pp.61-65
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• 2005

The objective of this work was to investigate the nutritional requirements for the growth of Actinomyces sp. 9RCC5 isolated from the rumen of a native goat in Korea. The growth of strain 9RCC5 on the basal medium or the medium minus certain ingredients from the basal medium demonstrated that strain 9RCC5 showed absolute requirement of vitamin B complex mixture, while hemin and volatile fatty acids (VFA) were stimulatory to growth to some extent. The 9RCC5 strain grew well with casein hydrolysate as the sole added nitrogen source. However, neither a complex of 18 amino acids nor ammonium sulfate effectively replaced casein hydrolysate. Vitamins such as riboflavin and pantothenate were essential for growth, while thiamin and biotin were stimulatory. With regard to VFA, the growth was stimulated by acetic acid but inhibited by valeric acid. Relatively large quantities of $Na^+$, $K^+$ and $Ca^{2+}$ were absolutely required for growth. Supplementation of clarified rumen fluid to the basal medium in a range of 0-10% (vol/vol) resulted in an increased rate of growth as well as an increased extent of growth.

• Animal cells and systems
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• 제1권4호
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• pp.589-594
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• 1997

The neuronal voltage-sensitive calcium channels (VSCCs) are multisubunit complexes consisting of $\alpha_1,\;\alpha_2-\delta$ and $\beta$ subunits. Heterologous expression and biochemical studies have shown that the activity of VSCCs is regulated by their $\beta$ subunits in a $\beta$ subunit isoform-specific manner. To elucidate the $\beta$ subunit identity of the P/Q-type calcium channel encoded by an $\alpha_{1A}$ subunit, which is exclusively expressed in the Purkinje and granule cell of the cerebellum, we have examined the spatial and temporal expression patterns of $\beta$ subunits and compared them with those of $\alpha_{1A}$ subunit in the developing rat cerebellum. Reverse transcriptase- polymerase chain reaction (RT-PCR) and Northern blot analysis have shown that $\beta_4$ subunit mRNA was prominently expressed in the cerebellum and much more abundant than any other distinct $\beta$ subunits. RNase protection assay has further demonstrated that the expression of $\alpha_{1A}$ and $\beta_4$ subunits increased during cerebellar development, while the amount of $\beta_2$ and $\beta_3$ mRNAs did not significantly change. In addition, a $\beta_4$ transcript was present in cultured cerebellar granule cells, but not in astrocyte cells, and the level of $\beta_4$ mRNA expression increased gradually in vitro seen as in vivo. Based on the spatial and temporal expression patterns of $\beta_4$ subunit, we conclude that $\beta_4$ may predominantly associate, but probably not exclusively, with the $\alpha_{1A}$ subunit in rat cerebellar granule cells.