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Journal of Biochemistry

Feng Li, Fujun Jin, Yiliang Wang, Danlin Zheng, Junwei Liu, Zhen Zhang, Rongze Wang, Dong Dong, Kai Zheng, Yifei Wang
Heat shock protein 90 (Hsp90) has been identified as an essential host factor for the infection and replication of several viruses, including HSV-1. Recent works have clearly shown that Hsp90 plays a role in the early stages of HSV-1 infection, including nuclear import and DNA replication. However, the role of Hsp90 in the late stages of HSV-1 infection remains unclear. In this study, we found that Hsp90 was up-regulated during late viral infection. Treatment with the Hsp90 inhibitor AT-533 significantly decreased the intracellular and extracellular virus titer, and strongly inhibited nucleocapsid egress from the nucleus...
August 10, 2018: Journal of Biochemistry
Nanami Kawaguchi, Kimie Date, Yusuke Suzuki, Chihiro Tomita, Rina Naradate, Tomoko Higami, Kosuke Nakamura, Kyoko Aikawa, Haruko Ogawa
An active recombinant human pancreatic lipase (recHPL) was successfully prepared for the first time from the Escherichia coli expression system using short strep-tag II (ST II). The recHPL-ST II was solubilized using 8 M urea from E. coli lysate and purified on a Strep-Tactin-Sepharose column. After refolding by stepwise dialyses in the presence of glycerol and Ca2+ for 2 days followed by gel filtration, 1.8-6 mg of active recHPL-ST II was obtained from 1 L of culture. The recHPL was non-glycosylated, but showed almost equal specific activity, pH-dependency, and time-dependent stability compared to those of native porcine pancreatic lipase (PPL) at 37 °C...
August 8, 2018: Journal of Biochemistry
Hiroki Yamaguchi, Moemi Tatsumi, Kazutoshi Takahashi, Uno Tagami, Masayuki Sugiki, Tatsuki Kashiwagi, Masafumi Kameya, Seiji Okazaki, Toshimi Mizukoshi, Yasuhisa Asano
L-Tryptophan oxidase, VioA from Chromobacterium violaceum, which has a high substrate specificity for tryptophan, is useful for quantitative assay of tryptophan. However, stability of wild type VioA is not enough for its application in clinical or industrial use. To improve the thermal stability of the enzyme, we developed a VioA (C395A) mutant, with higher stability than wild type VioA. The VioA (C395A) exhibited similar specificity and kinetic parameter for tryptophan to wild type. Conventionally, the quantity of tryptophan is determined by instrumental methods, such as high-performance liquid chromatography (HPLC) after pre-column-derivatization...
July 24, 2018: Journal of Biochemistry
Fumihiro Motojima, Katsuya Fujii, Masasuke Yoshida
Chaperonins assist folding of many cellular proteins, including essential proteins for cell viability. However, it remains unclear how chaperonin-assisted folding is different from spontaneous folding. Chaperonin GroEL/GroES facilitates folding of denatured protein encapsulated in its central cage but the denatured protein often escapes from the cage to the outside during reaction. Here, we show evidence that the in-cage-folding and the escape occur diverging from the same intermediate complex in which polypeptide is tethered loosely to the cage and partly protrudes out of the cage...
July 23, 2018: Journal of Biochemistry
Yu-Ichiro Nakajima
Polarized epithelia are a foundation of organ and appendage structures throughout Metazoa and serve as a physical barrier to preserve physiological functions. In proliferating epithelia, planar cell division occurs by orienting the mitotic spindle within the plane of the epithelium to ensure tissue organization. Conversely, loss of tissue architecture is a hallmark of carcinoma, and aberrant spindle orientation is hypothesized to contribute to tissue disorganization through dysplasia and cell dissemination...
July 18, 2018: Journal of Biochemistry
Shikshya Timalsina, Kyoko Arimoto-Matsuzaki, Masami Kitamura, Xiaoyin Xu, Qiu Wenzhe, Mari Ishigami-Yuasa, Hiroyuki Kagechika, Yutaka Hata
In eukaryotic cells, when exposed to certain types of stress including hypoxia, eIF2α is phosphorylated by several kinases including protein kinase R (PKR) and PKR-like endoplasmic reticulum kinase (PERK). Subsequently, protein translation is stopped and stress granules (SGs) are formed. Cancer cells form SGs under hypoxia. SGs accumulate apoptosis-related molecules and play anti-apoptotic roles. Thus, hypoxia-induced SG formation contributes to drug resistance in cancer cells. For this reason, inhibition of SG formation is expected to be beneficial in cancer therapy...
July 17, 2018: Journal of Biochemistry
Sofiane Hamidi, Guojun Sheng
Epithelial-Mesenchymal Transition (EMT) is a morphogenetic process of cells that adopt an epithelial organization in their developmental ontogeny or homeostatic maintenance. Abnormalities in EMT regulation result in many malignant tumors in the human body. Tumors associated with the hematopoietic system, however, are traditionally not considered to involve EMT and hematopoietic stem cells (HSCs) are generally not associated with epithelial characteristics. In this review, we discuss the ontogeny and homeostasis of adult HSCs in the context of EMT intermediate states...
July 17, 2018: Journal of Biochemistry
Hidetoshi Saze
Genomes of animals and plants contain a large number of transposable elements (TEs). TEs often transpose into genic regions, affecting expression of surrounding genes. Intragenic TEs mostly reside in introns, and in much the same way as intergenic TEs, they are targeted by repressive epigenetic marks for transcriptional silencing. Silenced intragenic TEs generally co-repress expression of associated genes, while in some cases they significantly enhance splicing and transcript elongation. Genomes have evolved molecular mechanisms that allow the presence of silenced TEs within transcriptionally permissive chromatin environments...
July 14, 2018: Journal of Biochemistry
Lingang Cui, Qingjun Meng, Jianguo Wen, Zechen Yan, Zhan Gao, Yudong Tian, Pengchao Xu, Pengchao Lian, Haizhou Yu
This study aimed to investigate the exact regulatory mechanisms of GRIM-19 in renal carcinoma. Tumor tissue samples from patients with renal carcinoma (n = 30, there were 7 cases of stage I, 8 cases of stage II, 8 cases of stage III, 7 cases of stage IV) and control subjects were selected from adjacent normal tissue (n = 10). Real-time quantitative PCR (qRT-PCR) and Western blotting was used to assess the level of GRIM-19, STAT3 and its downstream molecules. CD31 was detected by immunohistochemistry...
June 29, 2018: Journal of Biochemistry
Susumu Itoh, Fumiko Itoh
The TMEPAI family, composed of TMEPAI and C18ORF1, is known to inhibit TGF-β (transforming growth factor-β) signaling via its competition for binding of R-Smad (receptor-regulated Smad) with SARA (Smad anchor for receptor activation). However, TMEPAI has also been reported to be involved in AR (androgen receptor) signaling, PTEN (phosphatase and tensin homolog deleted on chromosome 10) signaling, and formation of autophagosomes in addition to degradation of TβRI (TGF-β type I receptor) through lysosomes...
June 26, 2018: Journal of Biochemistry
Takehiro Yasukawa, Dongchon Kang
While the majority of DNA is enclosed within the nucleus, the mitochondria also contain their own, separate DNA, the mitochondrial DNA (mtDNA). Mutations in mtDNA are associated with various human diseases, demonstrating the importance of mtDNA. Intensive studies over the last 18 years have demonstrated the presence of two distinct classes of mtDNA replication intermediates in mammals. One involves leading-strand DNA synthesis in the absence of synchronous lagging-strand DNA synthesis. Currently there are competing models in which the lagging-strand template is either systematically hybridized to processed mitochondrial transcripts, or coated with protein, until the lagging-strand DNA synthesis takes place...
June 20, 2018: Journal of Biochemistry
Abhijit Chowdhury, Teru Ogura, Masatoshi Esaki
Mitochondria continuously undergo coordinated fusion and fission during vegetative growth to keep their homogeneity and to remove damaged components. A cytosolic AAA ATPase, Cdc48, is implicated in the mitochondrial fusion event and turnover of a fusion-responsible GTPase in the mitochondrial outer membrane, Fzo1, suggesting a possible linkage of mitochondrial fusion and Fzo1 turnover. Here, we identified two Cdc48 cofactor proteins, Ubp3 and Ubx2, involving mitochondria regulation. In the absence of UBP3, mitochondrial fragmentation and aggregation were observed...
June 19, 2018: Journal of Biochemistry
Jung-Wei Chang, Yusuke Sato, Tetsuhiro Ogawa, Takatoshi Arakawa, Shuya Fukai, Shinya Fushinobu, Haruhiko Masaki
Colicins are protein toxins produced by and toxic to Escherichia coli strains. Colicin D consists of an N-terminal domain (NTD), central domain (CD), and C-terminal RNase domain (CRD). The cognate immunity protein, ImmD, is co-synthesized in producer cells to block the toxic tRNase activity of the CRD. Previous studies have reported the crystal structure of CRD/ImmD complex. Colicin D hijacks the surface receptor FepA and the energy transducer TonB system using the NTD for translocation across the outer membrane of the target cells...
June 14, 2018: Journal of Biochemistry
Yuan Li, Anna Rita Cappello, Luigina Muto, Emanuela Martello, Marianna Madeo, Rosita Curcio, Paola Lunetti, Susanna Raho, Francesco Zaffino, Luca Frattaruolo, Rosamaria Lappano, Rocco Malivindi, Marcello Maggiolini, Donatella Aiello, Carmela Piazzolla, Loredana Capobianco, Giuseppe Fiermonte, Vincenza Dolce
Several ATP-depending reactions take place in the endoplasmic reticulum (ER). Although in Saccharomyces cerevisiae ER the existence of a Sac1p-dependent ATP transport system was already known, its direct involvement in ATP transport was excluded. Here we report an extensive biochemical characterization of a partially purified adenine nucleotide transport system (ANTS) not dependent on Sac1p. Highly purified ER membranes from the wild-type and Δsac1 yeast strains reconstituted into liposomes transported ATP with the same efficiency...
June 8, 2018: Journal of Biochemistry
Seaim Lwin Aye, Kei Fujiwara, Nobuhide Doi
Engineering and design of genetic circuit in living cell is critical in accessing the beneficial application of synthetic biology. Directed evolution can avoid the complicated rational design of such circuit by screening or selecting functional circuit from non-functional one. Here we proposed a positive-negative selection system for selecting a transcription factor that activates gene expression in response to arsenic in solution. First, we developed a whole cell biosensor for sensing arsenite in liquid using a regulator (ArsR) and a reporter (GFP), and evaluated its performance...
June 8, 2018: Journal of Biochemistry
Takuma Kozono, Kazuko Tadahira, Wataru Okumura, Nao Itai, Miwa Tamura-Nakano, Taeko Dohi, Takashi Tonozuka, Atsushi Nishikawa
Jaw1/LRMP is characterized as a type II integral membrane protein that is localized to endoplasmic reticulum (ER), however, its physiological functions have been poorly understood. An alignment of amino acid sequence of Jaw1 with KASH proteins, outer nuclear membrane proteins, revealed that Jaw1 has a partial homology to the KASH domain. Here, we show that the function of Jaw1 is to maintain nuclear shape in mouse melanoma cell line. The siRNA-mediated knockdown of Jaw1 caused a severe defect in nuclear shape, and the defect was rescued by ectopic expression of siRNA-resistant Jaw1...
June 6, 2018: Journal of Biochemistry
Kei Sadakane, Islam M D Alrazi, Shinsaku Maruta
Mitotic kinesin Eg5 plays an important physiological role in cell division. Several small-molecule inhibitors of Eg5 are the focus of cancer therapies. Azobenzene is a photochromic compound exhibiting cis-trans isomerization upon ultraviolet (UV) and visible (VIS) light irradiation. Photochromic compounds of azobenzene derivatives, mimicking Eg5 specific inhibitors of STLC, indicated photoreversible inhibitory effects on Eg5 ATPase activity; however, the photoreversible switching efficiency was not significant...
May 31, 2018: Journal of Biochemistry
Satoshi Inouye, Yuri Tomabechi, Takamitsu Hosoya, Shun-Ichi Sekine, Mikako Shirouzu
cf3-Aequorin is one of the semi-synthetic aequorins that was produced by replacing 2-peroxycoelenterazine (CTZ-OOH) in native aequorin with a 2-peroxycoelenterazine analog, and it was prepared using the C2-modified trifluoromethyl analog of coelenterazine (cf3-CTZ) and the histidine-tagged apoaequorin expressed in Escherichia coli cells. The purified cf3-aequorin showed a slow luminescence pattern with half-decay time of Imax of 5.0 s. This is much longer than that of 0.9 s for native aequorin, and its luminescence capacity was estimated to be 72...
May 23, 2018: Journal of Biochemistry
Kenichi Asano, Kenta Kikuchi, Masato Tanaka
Tissue macrophages comprise heterogeneous subsets that differ in localization, phenotype and ontogeny. They acquire tissue-specific phenotype in order to maintain normal tissue physiology. This review summarizes the current knowledge about the functions of CD169-positive macrophage subset residing in the lymphoid organs and intestinal tract. Strategically positioned at the interface between tissue and circulating fluid, CD169+ macrophages in the lymphoid organs capture blood- and lymph-borne particulate materials...
August 1, 2018: Journal of Biochemistry
Masafumi Noda, Sachiko Sugimoto, Ikue Hayashi, Narandalai Danshiitsoodol, Mitsuhiro Fukamachi, Masanori Sugiyama
A lactic acid bacterium Lactobacillus paracasei IJH-SONE68, which was isolated from a fig leaf by our group, was found to produce both acidic and neutral exopolysaccharides (EPSs). The nuclear magnetic resonance analysis demonstrates that the former EPS is composed primarily of mannose, and the latter one consists of the α-1, 6-linked glycan chains made of N-acetylglucosamine (GlcNAc). The presence of α-1, 6-linked GlcNAc polysaccharide is first reported in prokaryotes. Furthermore, to reveal the EPS-biosynthetic gene organization in the IJH-SONE68 strain, in the present study, we determined the whole-genome sequence...
August 1, 2018: Journal of Biochemistry
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