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Journal of Biochemistry

Akihiko Kanou, Koichiro Kako, Keiko Hirota, Akiyoshi Fukamizu
No abstract text is available yet for this article.
February 19, 2018: Journal of Biochemistry
Zhao Peng, Miyu Takeshita, Nao Shibata, Hideaki Tada, Yoshikazu Tanaka, Jun Kaneko
Staphylococcus aureus bi-component pore-forming toxins consist of S- and F-components, and form hetero-octameric beta-barrel pores on target blood cell membranes. Among them, γ-hemolysin (Hlg2 and LukF) and LukED (LukE and LukD) possess hemolytic activity, whereas the Panton -Valentine leukocidin (LukS-PV and LukF-PV) does not lyse human erythrocytes.Here, we focused on four loop structures in the rim domain of S-component, namely loops -1, -2, -3, and -4, and found that replacement of loop-4 in both Hlg2 and LukE with that of LukS-PV abolished their hemolytic activity...
February 17, 2018: Journal of Biochemistry
Akshari Gupta, Daiju Kitagawa
Several decades of centriole research have revealed the beautiful symmetry present in these microtubule-based organelles, which are required to form centrosomes, cilia, and flagella in many eukaryotes. Centriole architecture is largely conserved across most organisms, however, individual centriolar features such as the central cartwheel or microtubule walls exhibit considerable variability when examined with finer resolution. Here, we review the ultrastructural characteristics of centrioles in commonly studied organisms, highlighting the subtle and not-so-subtle differences between specific structural components of these centrioles...
February 16, 2018: Journal of Biochemistry
Fengying Tang, Megan S Lord, William B Stallcup, John M Whitelock
Chondroitin sulfate proteoglycan 4 (CSPG4) is a cell surface proteoglycan highly expressed by tumor, perivascular and oligodendrocyte cells and known to be involved cell adhesion and migration. This study showed that CSPG4 was present as a proteoglycan on the cell surface of two melanoma cell lines, MM200 and Me1007, as well as shed into the conditioned medium. CSPG4 from the two melanoma cell lines differed in the amount of chondroitin sulfate (CS) decoration, as well as the way the protein core was fragmented...
February 15, 2018: Journal of Biochemistry
Daisuke Kato, Kei Eto, Junichi Nabekura, Hiroaki Wake
Electrical activity is essential for brain function. However, neurons, the electrically active cells, are less numerous than the non-electrical glial cells in the central nervous system. The non-electrical components modify the function of neural circuits, depending on the electrical neuronal activity, by wrapping synapses, myelinating axons, and phagocytozing the neuronal components. Moreover, recent evidence has suggested that they contribute to neurological and psychiatric disease by regulating neuronal circuits, ultimately affecting their behavior...
February 13, 2018: Journal of Biochemistry
Ziaur Rahman, Yuta Tsujimori, Megumi Maeda, Anowar Hossain, Takeshi Ishimizu, Yoshinobu Kimura
In a previous study, we molecular-characterized a tomato (Solanum lycopersicum) α1,3/4-fucosidase (α-Fuc'ase S1-1) encoded in a tomato gene (Solyc03g006980), indicating that α-Fuc'ase S1-1 is involved in the turnover of Lea epitope-containing N-glycans. In this study, we have characterized another tomato gene (Solyc11g069010) encoding α1,3/4-fucosidase (α-Fuc'ase S1-2), which is also active toward the complex type N-glycans containing Lea epitope(s). The baculovirus-insect cell expression system was used to express that α-Fuc'ase S1-2 with anti-FLAG tag, and the expression product (rFuc'ase Sl-2), was found as a 65 kDa protein using SDS-PAGE and has an optimum pH of around 5...
February 10, 2018: Journal of Biochemistry
Teisuke Takita, Jun Qian, Hongmin Geng, Zejian He, Sho Nemoto, Mariko Mori, Keisuke Tanaka, Shunji Hattori, Kenji Kojima, Kiyoshi Yasukawa
The collagenase produced by a gram-negative bacterium Grimontia hollisae strain 1706B (Ghcol) degrades collagen more efficiently than that produced by a gram-positive bacterium Clostridium histolyticum (Chcol), which is currently the most widely used collagenase in industry (Teramura et al. (2011) J. Bacteriol.193, 3049‒3056). Here, we compared the Ghcol and Chcol activities using two synthetic substrates. In the hydrolysis of (7-methoxycoumarin-4-yl)acetyl-L-Lys-L-Pro-L-Leu-Gly-L-Leu-[N3-(2,4-dinitrophen-yl)-L-2,3-diaminopropioyl]-L-Ala-L-Arg-NH2, Ghcol exhibited 350-fold higher activity than Chcol in the absence of CaCl2 and NaCl...
February 10, 2018: Journal of Biochemistry
Daisuke Saigusa, Norio Suzuki, Yotaro Matsumoto, Keiko Umeda, Yoshihisa Tomioka, Seizo Koshiba, Masayuki Yamamoto
Roxadustat (FG-4592, Rox) is a stabilizer for hypoxia-inducible transcription factors (HIFs), which induce production of the erythroid growth factor erythropoietin, and has been listed by the World Anti-Doping Agency as a prohibited substance for athletes since 2011. Although the detection technologies for Rox and its glucuronide-conjugated metabolite (Rox-Gluc) have been developed exploiting triple quadrupole mass spectrometry (MS/MS), the production of metabolites from Rox in the human body remains to be clarified...
February 9, 2018: Journal of Biochemistry
Kouki Tadai, Tatsumasa Shioiri, Jun Tsuchimoto, Naoko Nagai, Hideto Watanabe, Nobuo Sugiura
Receptor type of protein tyrosine phosphatase sigma (RPTPσ) functions as a glycosaminoglycan (GAG) receptor of neuronal cells in both the central and peripheral nervous systems. Both chondroitin sulfate (CS) and heparan sulfate (HS) are important constituents of GAG ligands for RPTPσ, although they have opposite effects on neuronal cells. CS inhibits neurite outgrowth and neural regeneration through RPTPσ, whereas HS enhances them. We prepared recombinant RPTPσ N-terminal fragment containing the GAG binding site and various types of biotin-conjugated GAG (CS and HS) with chemical modification and chemo-enzymatic synthesis...
February 6, 2018: Journal of Biochemistry
Mitsuhiro Itaya, Hiroko Kusakabe, Mitsuru Sato, Masaru Tomita, Rintaro Sato
Synechococcus elongatus PCC7942, a cyanobacterium that uses light and carbon dioxide to grow, has a high ability to incorporate DNA by transformation. To assess the effective delivery of large DNA in plasmid form, we cloned the endogenous plasmid pANL (46.4 kbp) into a BAC vector of Escherichia coli. The plasmid p38ANL (54.3 kbp) replaced the native plasmid. To assess the delivery of larger DNA into PCC7942, p38ANL was fused to the broad-host-range conjugal transfer plasmid pUB307IP (53.5 kbp). The resulting plasmid pUB307IP501 (107...
February 6, 2018: Journal of Biochemistry
Yuriko Egashira, Satoru Nagatoishi, Masato Kiyoshi, Akiko Ishii-Watabe, Kouhei Tsumoto
Silkworm-baculovirus expression systems are efficient means for production of recombinant proteins that provide high expression levels and post-translational modifications. Here, we characterized the stability, glycosylation pattern, and antibody-dependent cell-mediated cytotoxicity activity of anti-HER2 monoclonal antibodies containing native or glycoengineered mammalian-like N-glycans that were produced by using a silkworm-baculovirus expression system. Compared with a monoclonal antibody produced by using a Chinese hamster ovary cell expression system, the glycoengineered monoclonal antibody had comparable thermal stability and a higher antibody-dependent cell-mediated cytotoxicity activity...
February 5, 2018: Journal of Biochemistry
Takeshi Kaizuka, Toru Takumi
Synaptic connections are essential for neural circuits in order to convey brain functions. The postsynaptic density (PSD) is a huge protein complex associated with postsynaptic membranes of excitatory synapses. In mammals, the PSD is composed of more than 1,000 proteins including receptors, scaffold proteins, signaling enzymes, and cytoskeletal proteins. PSD proteins are crucial for synaptic transmission and plasticity. Proteomic studies have revealed the composition of PSD proteins in various species, brain regions, and specific physiological conditions...
February 5, 2018: Journal of Biochemistry
Takuya Ogawa, Asako Tanaka, Jun Kawamoto, Tatsuo Kurihara
1-Acyl-sn-glycerol-3-phosphate acyltransferase (designated as PlsC in bacteria) catalyzes the acylation of lysophosphatidic acid and is responsible for the de novo production of phosphatidic acid, a precursor for the synthesis of various membrane glycerophospholipids. Because PlsC is an integral membrane protein, it is generally difficult to solubilize it without causing its inactivation, which has been hampering its biochemical characterization despite its ubiquitous presence and physiological importance. Most biochemical studies of PlsC have been carried out by using crude membrane preparations or intact cells...
February 5, 2018: Journal of Biochemistry
Kei-Ichi Yamaguchi, Ryo P Honda, Abdelazim Elsayed Elhelaly, Kazuo Kuwata
Although pulsatile irradiation of ultrasonication is frequently used for generating amyloid fibrils in vitro, the potential for inducing amyloid fibrillation of proteins during continuous ultrasonication is unknown. In this study, we implemented a continuous irradiation system and measured far-ultraviolet circular dichroism in a real-time manner. During the continuous ultrasonication, the conformation of full-length mouse prion protein (mPrP) was rapidly altered without a lag time, and electron microscopy revealed that distorted fibrils, β-oligomers, and amorphous aggregates were formed at pH 2...
February 2, 2018: Journal of Biochemistry
So-Ichiro Fukada
Skeletal muscle is composed of multinuclear cells called myofibers. Muscular dystrophy (a genetic muscle disorder) induces instability in the cell membrane of myofibers and eventually causes myofiber damage. Non-genetic muscle disorders, including sarcopenia, diabetes, bedridden immobility, and cancer cachexia, lead to atrophy of myofibers. In contrast, resistance training induces myofiber hypertrophy. Thus, myofibers exhibit a plasticity that is strongly affected by both intrinsic and extrinsic factors. There is no doubt that muscle stem cells (MuSCs, also known as muscle satellite cells) are indispensable for muscle repair/regeneration, but their contributions to atrophy and hypertrophy are still controversial...
January 31, 2018: Journal of Biochemistry
Motoi Kanagawa, Tatsushi Toda
Glycosylation is a crucial posttranslational modification that is involved in numerous biological events. Therefore, abnormal glycosylation can impair the functions of glycoproteins or glycolipids and is occasionally associated with cell dysfunction and human diseases. For example, aberrant glycosylation of dystroglycan, a cellular receptor for matrix and synaptic proteins, is associated with muscular dystrophy and lissencephaly. Dystroglycan sugar chains are required for high-affinity binding to ligand proteins, and thus disruption of dystroglycan-ligand linkages underlies disease conditions...
January 31, 2018: Journal of Biochemistry
Wataru Yokoyama, Keiko Hirota, Huahua Wan, Naoaki Sumi, Mai Miyata, Sho Araoi, Naoto Nomura, Koichiro Kako, Akiyoshi Fukamizu
RNAs are post-transcriptionally modified in all kingdoms of life. Of these modifications, base methylations are highly conserved in eukaryote ribosomal RNA (rRNA). Recently, rRNA processing protein 8 (Rrp8) and nucleomethylin (NML) were identified as factors of N1-methyladenosine (m1A) modification in yeast 25S and mammalian 28S rRNA, respectively. However, m1A modification of rRNA is still poorly understood in Caenorhabditis elegans. Here, using the liquid chromatography/tandem mass spectrometry analysis and RNA immunoprecipitation (RIP) assay, we have identified that m1A modification is located around position 674 (A674) of 26S rRNA in C...
January 27, 2018: Journal of Biochemistry
Tomotaka Nagasawa, Hikaru Nakamichi, Yoichiro Hama, Shigeki Higashiyama, Yasuyuki Igarashi, Susumu Mitsutake
GPR120 is a receptor for long chain fatty acids and is expressed in small intestinal endocrine cells, L cells and adipose tissue. Activation of GPR120 promotes the secretion of incretin GLP-1, which is known to have effects on anti-metabolic syndrome. As such, GPR120 is a potential target of pharmaceuticals for type II diabetes. In this study, we performed ligand-screening for GPR120 on glycero- and sphingo-type lipids and their derivatives using a TGFα-shedding assay. We found that phytosphingosine (PHS) activates GPR120 in a manner comparable to the natural ligand α-linolenic acid (ALA) and superior to that of the synthetic ligand GW9508...
January 24, 2018: Journal of Biochemistry
Yukio Nisimoto, Hisamitsu Ogawa, Yuzo Kadokawa, Shanlou Qiao
Nox4, a member of the NADPH- and oxygen-dependent oxidoreductases that generate reactive oxygen species (ROS), is widely expressed and constitutively active. To understand better its function and regulation, specific mutations in the Nox4 dehydrogenase (DH) domain were examined for effects on Nox4 oxidase activity. Transfection of His6-tagged Nox4 increased the amount of p22phox subunit in HEK293 cells, and a higher level of the heterodimer was observed in the nucleus-enriched fraction (NEF). NEF from Nox4-expressing HEK293 cells exhibited oxygen and H2O2 concentration-dependent NADPH oxidation rate...
January 22, 2018: Journal of Biochemistry
Ryota Saito, Yutaro Saito, Hikaru Nakazawa, Takamitsu Hattori, Izumi Kumagai, Mitsuo Umetsu, Koki Makabe
Biomolecules which recognize inorganic materials and metal surfaces gain much attention for creating new type of nanomaterials and sensors. 4F2, a camelid VHH antibody, recognizes ZnO surface and has been applied for sensor applications. 4F2 was constructed sequential CDR replacement on the parental VHH antibody, termed the Construction of Antibody by Integrating Grafting and Evolution Technology; CAnIGET procedure. Here, we evaluate the influence of CDR replacements during 4F2 generation using calorimetric technique...
January 22, 2018: Journal of Biochemistry
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