22 results on '"Sugiyama, Kanako"'
Search Results
2. Successful Bridge-to-Recovery Treatment in a Young Patient with Fulminant Eosinophilic Myocarditis: Roles of a Percutaneous Ventricular Assist Device and Endomyocardial Biopsy
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Hasegawa-Tamba, Saki, Sugi, Keiki, Gatate, Yodo, Sugiyama, Kanako, Muramatsu, Toshihiro, Nishimura, Shigeyuki, Yasuda, Masanori, Fukushima, Kenji, and Nakano, Shintaro
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Article Subject - Abstract
Eosinophilic myocarditis (EM) is a rare condition characterized by myocardial eosinophilic infiltration due to various underlying etiologies. The patient with EM may benefit from appropriate use of mechanical circulatory support (MCS) that acts as a bridge to myocardial recovery in response to effective immunosuppressive therapy. A 16-year-old boy presented with cardiogenic shock due to fulminant myocarditis, for which a percutaneous ventricular assist device (PVAD) was immediately inserted. Based on the histological diagnosis of EM, immunosuppressive therapy was immediately commenced, leading to improvement of left-ventricular ejection fraction (27% to 47%). The PVAD was successfully removed on day 7. Cardiac magnetic resonance imaging and dual-tracer myocardial scintigraphy suggested limited extent of irreversible myocardial damage. For fulminant EM, the short-term use of PVAD, together with immunosuppressive therapy guided by an immediate histological investigation, may be an effective bridging strategy to myocardial recovery.
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- 2019
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3. Interstitial pneumonia caused by inhalation of fumes of nickel and chrome
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HISATOMI, KEIKO, ISHII, HIROSHI, HASHIGUCHI, KOJI, SEKI, MASAFUMI, IDE, MIOKO, SUGIYAMA, KANAKO, ISHIMOTO, HIROSHI, NAKAYAMA, SEIKO, Mukae, HIROSHI, and Kohno, SHIGERU
- Published
- 2006
4. Elevated levels of interferon γ-inducible protein-10 and epithelial neutrophil-activating peptide-78 in patients with pulmonary sarcoidosis
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SUGIYAMA, KANAKO, MUKAE, HIROSHI, ISHII, HIROSHI, KAKUGAWA, TOMOYUKI, ISHIMOTO, HIROSHI, NAKAYAMA, SEIKO, SHIRAI, RYO, FUJII, TAKESHI, MIZUTA, YOHEI, and KOHNO, SHIGERU
- Published
- 2006
5. Expression of HSP47 in Usual Interstitial Pneumonia and Nonspecific Interstitial Pneumonia
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Mine Mariko, Sugiyama Kanako, Yoshioka Sumako, Sakamoto Noriho, Nakayama Seiko, Ishii Hiroshi, Hayashi Tomayoshi, Mukae Hiroshi, Kakugawa Tomoyuki, Mizuta Yohei, and Kohno Shigeru
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usual interstitial pneumonia ,nonspecific interstitial pneumonia ,collagen vascular disease ,heat shock protein 47 ,type I procollagens ,Diseases of the respiratory system ,RC705-779 - Abstract
Abstract Background Heat shock protein (HSP) 47, a collagen-specific molecular chaperone, is involved in the processing and/or secretion of procollagens, and its expression is increased in various fibrotic diseases. The aim of this study was to determine whether quantitative immunohistochemical evaluation of the expression levels of HSP47, type I procollagen and α-smooth muscle actin (SMA) allows the differentiation of idiopathic usual interstitial pneumonia (UIP) from UIP associated with collagen vascular disease (CVD) and idiopathic nonspecific interstitial pneumonia (NSIP). Methods We reviewed surgical lung biopsy specimens of 19 patients with idiopathic UIP, 7 with CVD-associated UIP and 16 with idiopathic NSIP and assigned a score for the expression of HSP47, type I procollagen and α-SMA in type II pneumocytes and/or lung fibroblasts (score 0 = no; 1 = weak; 2 = moderate; 3 = strong staining). Results The expression level of HSP47 in type II pneumocytes of idiopathic UIP was significantly higher than in CVD-associated UIP and idiopathic NSIP. The expression of HSP47 in fibroblasts was significantly higher in idiopathic UIP and idiopathic NSIP than in CVD-associated UIP. The expression of type I procollagen in type II pneumocytes was significantly higher in idiopathic UIP than in idiopathic NSIP. The expression of type I procollagen in fibroblasts was not different in the three groups, while the expression of α-SMA in fibroblasts was significantly higher in idiopathic UIP than in idiopathic NSIP. Conclusion Our results suggest the existence of different fibrotic pathways among these groups involved in the expression of HSP47 and type I procollagen.
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- 2005
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6. Spontaneous Pneumothorax and Pneumomediastinum in Patients with Idiopathic Interstitial Pneumonias
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Sakamoto, Noriho, Mukae, Hiroshi, Ishii, Hiroshi, Sugiyama, Kanako, Kakugawa, Tomoyuki, Ishimoto, Hiroshi, Yoshioka, Sumako, Nakayama, Seiko, Abe, Koh, Fujii, Takeshi, and Kohno, Shigeru
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Steroidtreatment ,Idiopathicpulmonaryfibrosis ,Pneumomediastinum ,Pneumothorax ,respiratory system ,Idiopathicinterstitialpneumonia ,humanities ,respiratory tract diseases - Abstract
Pneumothorax and pneumomediastinum sometimes occur in patients with idiopathic interstitial pneumonias (IIPs). Although steroid treatment is commonly used for IIPs, some reports indicated that this treatment could cause pneumothorax (PT) or pneumomediastinum (PM) in patients with IIPs. The aim of the present study was to evaluate the clinical features of PT and PM in patients with IIPs, and to assess their association with steroid treatment. We enrolled 77 patients with IIPs and divided them into four groups: (1) idiopathic pulmonary fibrosis (IPF) patients without PT and PM [IPF PT/PM(-) group, n = 38]; (2) IPF patients with PT and/or PM [IPF PT/PM(+) group, n = 6]; (3) non-IPF patients without these complications [non-IPF PT/PM(-) group, n = 29]; and (4) non-IPF patients with PT and/or PM [non-IPF PT/PM(+) group, n = 4]. We reviewed and compared the clinical, radiological and laboratory findings among the four groups. We also focused on the above 10 IIPs patients with PT and/or PM to describe the details of their clinical features. Resultingly, IPF PT/PM(+) and non-IPF PT/PM(+) groups showed low percentage of vital capacity (%VC) compared with IPF PT/PM(-) and non-IPF PT/PM(-) groups, respectively. Six of the 10 cases with PT/PM were treated with steroids about 2 weeks before the development of PT and/or PM. We concluded that PT and PM could arise in patients with IIPs, especially in cases with severe restrictive ventilatory impairment. Our results further suggest that clinicians should be aware of these complications after starting steroid treatment for interstitial lung diseases., Acta medica Nagasakiensia. 2006, 51(1), p.23-26
- Published
- 2006
7. Structure-based analysis of domain function of chitin oligosaccharide deacetylase from Vibrio parahaemolyticus.
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Hirano, Takako, Sugiyama, Kanako, Sakaki, Yuta, Hakamata, Wataru, Park, Sam-Yong, and Nishio, Toshiyuki
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CHITIN deacetylase , *OLIGOSACCHARIDES , *VIBRIO parahaemolyticus , *CRYSTAL structure , *CARBOHYDRATES , *MATHEMATICAL domains - Abstract
The X-ray crystal structure of chitin oligosaccharide deacetylase from Vibrio parahaemolyticus ( Vp -COD) was determined at an 1.35 Å resolution. The amino acid sequence and structure of Vp -COD show that the enzyme comprises one polysaccharide deacetylase domain (PDD) and two carbohydrate-binding domains (CBDs). On the basis of a chitin-binding assay with Vp -COD and its CBDs-deleted mutant, it was confirmed that CBDs can adhere to chitin. The catalytic activity of the CBDs-deleted mutant was only mildly depressed compared with that of Vp -COD, indicating that CBDs are unlikely to affect the configuration of the active center residues in active site of PDD. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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8. Capturing the Hemoglobin Allosteric Transition in a Single Crystal Form.
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Shibayama, Naoya, Sugiyama, Kanako, Tame, Jeremy R. H., and Sam-Yong Park
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HEMOGLOBINS , *SINGLE crystals , *ALLOSTERIC regulation , *PROTEINS , *X-ray diffraction - Abstract
Allostery in many oligomeric proteins has been postulated to occur via a ligand-binding-driven conformational transition from the tense (T) to relaxed (R) state, largely on the basis of the knowledge of the structure and function of hemoglobin, the most thoroughly studied of all allosteric proteins. However, a growing body of evidence suggests that hemoglobin possesses a variety of intermediates between the two end states. As such intermediate forms coexist with the end states in dynamic equilibrium and cannot be individually characterized by conventional techniques, very little is known about their properties and functions. Here we present complete structural and functional snapshots of nine equilibrium conformers of human hemoglobin in the half-liganded and fully liganded states by using a novel combination of X-ray diffraction analysis and microspectrophotometric O2 equilibrium measurements on three isomorphous crystals, each capturing three distinct equilibrium conformers. Notably, the conformational set of this crystal form varies according to shifts in the allosteric equilibrium, reflecting the differences in hemoglobin ligation state and crystallization solution conditions. We find that nine snapshot structures cover the complete conformational space of hemoglobin, ranging from T to R2 (the second relaxed quaternary structure) through R, with various relaxed intermediate forms between R and R2. Moreover, we find a previously unidentified intermediate conformer, between T and R, with an intermediate O2 affinity, sought by many research groups over a period of decades. These findings reveal a comprehensive picture of the equilibrium conformers and transition pathway for human hemoglobin. [ABSTRACT FROM AUTHOR]
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- 2014
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9. Homopiperazine Derivatives as a Novel Class of Proteasome Inhibitors with a Unique Mode of Proteasome Binding.
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Kikuchi, Jiro, Shibayama, Naoya, Yamada, Satoshi, Wada, Taeko, Nobuyoshi, Masaharu, Izumi, Tohru, Akutsu, Miyuki, Kano, Yasuhiko, Sugiyama, Kanako, Ohki, Mio, Park, Sam-Yong, and Furukawa, Yusuke
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PROTEASOME inhibitors ,PIPERAZINE ,HOMEOSTASIS ,MULTIPLE myeloma treatment ,INTRAVENOUS injections ,DRUG resistance ,APOPTOSIS ,ONCOLOGY - Abstract
The proteasome is a proteolytic machinery that executes the degradation of polyubiquitinated proteins to maintain cellular homeostasis. Proteasome inhibition is a unique and effective way to kill cancer cells because they are sensitive to proteotoxic stress. Indeed, the proteasome inhibitor bortezomib is now indispensable for the treatment of multiple myeloma and other intractable malignancies, but is associated with patient inconvenience due to intravenous injection and emerging drug resistance. To resolve these problems, we attempted to develop orally bioavailable proteasome inhibitors with distinct mechanisms of action and identified homopiperazine derivatives (HPDs) as promising candidates. Biochemical and crystallographic studies revealed that some HPDs inhibit all three catalytic subunits (ß 1, ß 2 and ß 5) of the proteasome by direct binding, whereas bortezomib and other proteasome inhibitors mainly act on the ß5 subunit. Proteasome-inhibitory HPDs exhibited cytotoxic effects on cell lines from various hematological malignancies including myeloma. Furthermore, K-7174, one of the HPDs, was able to inhibit the growth of bortezomib-resistant myeloma cells carrying a ß5-subunit mutation. Finally, K-7174 had additive effects with bortezomib on proteasome inhibition and apoptosis induction in myeloma cells. Taken together, HPDs could be a new class of proteasome inhibitors, which compensate for the weak points of conventional ones and overcome the resistance to bortezomib. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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10. Structures and Oxygen Affinities of Crystalline Human Hemoglobin C(β6 Glu → Lys) in the Rand R2 Quaternary Structu res*.
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Shibayama, Naoya, Sugiyama, Kanako, and Sam-Yong Park
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Recent crystallographic studies suggested that fully liganded human hemoglobin can adopt multiple quaternary conformations that include the two previously solved relaxed conformations, R and R2, whereas fully unliganded deoxyhemoglobin may adopt only one T (tense) quaternary conformation. An important unanswered question is whether R, R2, and other relaxed quaternary conformations represent different physiological states with different oxygen affinities. Here, we answer this question by showing the oxygen equilibrium curves of single crystals of human hemoglobin in the R and R2 state. In this study, we have used a naturally occurring mutant hemoglobin C (j36 GIu-~Lys) to stabilize the R and R2 crystals. Additionally, we have refined the x-ray crystal structure of carbonmonoxyhemoglobin C, in the R and R2 state, to 1.4 and 1.8 A resolution, respectively, to compare precisely the structures of both types of relaxed states. Despite the large quaternary structural difference between the R and R2 state, both crystals exhibit similar noncooperative oxygen equilibrium curves with a very high affinity for oxygen, comparable with the fourth oxygen equilibrium constant (K4) of human hemoglobin in solution. One small difference is that the R2 crystals have an oxygen affinity that is 2-3 times higher than that of the R crystals. These results demonstrate that the functional difference between the two typical relaxed quaternary conformations is small and physiologically less important, indicating that these relaxed conformations simply reflect a structural polymorphism of a high affinity relaxed state. [ABSTRACT FROM AUTHOR]
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- 2011
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11. Structural insight into the essential PB1–PB2 subunit contact of the influenza virus RNA polymerase.
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Sugiyama, Kanako, Obayashi, Eiji, Kawaguchi, Atsushi, Suzuki, Yukari, Tame, Jeremy R. H., Nagata, Kyosuke, and Park, Sam-Yong
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INFLUENZA viruses , *RNA polymerases , *ENZYME activation , *ENZYME regulation , *VIRAL genetics , *GENETICS - Abstract
Influenza virus RNA-dependent RNA polymerase is a multi-functional heterotrimer, which uses a ‘cap-snatching’ mechanism to produce viral mRNA. Host cell mRNA is cleaved to yield a cap-bearing oligonucleotide, which can be extended using viral genomic RNA as a template. The cap-binding and endonuclease activities are only activated once viral genomic RNA is bound. This requires signalling from the RNA-binding PB1 subunit to the cap-binding PB2 subunit, and the interface between these two subunits is essential for the polymerase activity. We have defined this interaction surface by protein crystallography and tested the effects of mutating contact residues on the function of the holo-enzyme. This novel interface is surprisingly small, yet, it has a crucial function in regulating the 250 kDa polymerase complex and is completely conserved among avian and human influenza viruses. [ABSTRACT FROM AUTHOR]
- Published
- 2009
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12. Crystal structure of the biotin carboxylase domain of human acetyl-CoA carboxylase 2.
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Cho, Yong Soon, Lee, Jae Il, Shin, Dongkyu, Kim, Hyun Tae, Cheon, Young Hoon, Seo, Chang Il, Kim, Yong Eun, Hyun, Young-Lan, Lee, Yoon Sup, Sugiyama, Kanako, Park, Sam-Yong, Ro, Seonggu, Cho, Joong Myung, Lee, Tae Gyu, and Heo, Yong-Seok
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- 2008
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13. Comparison of BALF concentrations of ENA-78 and IP10 in patients with idiopathic pulmonary fibrosis and nonspecific interstitial pneumonia.
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Nakayama, Seiko, Mukae, Hiroshi, Ishii, Hiroshi, Kakugawa, Tomoyuki, Sugiyama, Kanako, Sakamoto, Noriho, Fujii, Takeshi, Kadota, Jun-ichi, and Kohno, Shigeru
- Abstract
Summary: Background: Epithelial neutrophil-activating peptide 78 (ENA-78) and interferon -inducible protein 10 (IP10) belong to the CXC chemokine family and are considered to be important factors in idiopathic pulmonary fibrosis (IPF). Idiopathic nonspecific interstitial pneumonia (NSIP) and IPF are the two largest subsets of idiopathic interstitial pneumonias (IIP). In patients with NSIP, the prognosis is generally good compared with IPF. Therefore, the pathogenesis of NSIP seems to be different from that of IPF, but this remains unclear. The aim of the present study was to evaluate the contribution of ENA-78 and IP10 in the two diseases. Methods: We measured the levels of ENA-78 and IP10 in serum and bronchoalveolar lavage fluid (BALF) of patients with IPF (), idiopathic NSIP () and healthy subjects () by enzyme-linked immunosorbent assays. Results: The level of ENA-78 in BALF was significantly higher in IPF patients than in NSIP patients and controls. Serum levels of ENA-78 and BALF levels of IP10 in NSIP patients were significantly higher than in patients with IPF and controls. In BALF of patients with NSIP, IP10 level significantly correlated with the absolute number of lymphocytes. In IPF patients, BALF IP10 levels also correlated with the proportion of lymphocytes in BALF. Conclusion: Our results show distinct profiles of CXC chemokines in IPF and NSIP, and suggest that these chemokines play an important role in inflammatory cell recruitment into the lung in patients with IIP. [Copyright &y& Elsevier]
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- 2005
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14. Differential effects of α- and β-defensin on cytokine production by cultured human bronchial epithelial cells.
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Sakamoto, Noriho, Mukae, Hiroshi, Fujii, Takeshi, Ishii, Hiroshi, Yoshioka, Sumako, Kakugawa, Tomoyuki, Sugiyama, Kanako, Mizuta, Yohei, Jun-ichi Kadota, Nakazato, Masamitsu, and Kohno, Shigeru
- Subjects
CYTOKINES ,NEUTROPHILS ,PEPTIDES ,INTERLEUKIN-8 ,INTERLEUKINS ,EPITHELIAL cells ,CELLULAR immunity ,IMMUNOREGULATION - Abstract
Defensins are cysteine-rich cationic antimicrobial peptides that play an important role in innate immunity and are known to contribute to the regulation of host adaptive immunity. In addition to direct antimicrobial activities, it has been recently reported that α-defensins, mainly present in neutrophils in the lung, have a cytotoxic effect and induce IL-8 production from airway epithelial cells. Although β-defensins are expressed in epithelial cells in various tissues, including lung, there are no reports of their effects on cytokine synthesis in airway epithelial cells. The aim of the present study was to determine the effects of both α- and β-defensins on the cytokine production, transcription factor binding activity, and cytotoxicity in primary cultured human bronchial epithelial cells (HBECs). We used human neutrophil peptide-1 (HNP-1; α-defensin) and human β-defensin-2 (HBD-2) to stimulate HBECs. The results showed that treatment of HBECs with HNP-1, but not HBD-2, increased IL-8 and IL-1β mRNA expression in a dose-dependent manner and also enhanced IL-8 protein secretion and NF-κB DNA binding activity. The 24-h treatments with >20 μg/ml of HNP-1 or >50 μg/ml of HBD-2 were cytotoxic to HBECs. These results suggest that α- and β-defensins have different effects on cytokine synthesis by airway epithelial cells, and we speculate that they play different roles in inflammatory lung diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2005
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15. Expression of HSP47 in Usual Interstitial Pneumonia and Nonspecific Interstitial Pneumonia.
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Kakugawa, Tomoyuki, Mukae, Hiroshi, Hayashi, Tomayoshi, Ishii, Hiroshi, Nakayama, Seiko, Sakamoto, Noriho, Yoshioka, Sumako, Sugiyama, Kanako, Mine, Mariko, Mizuta, Yohei, and Kohno, Shigeru
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PULMONARY fibrosis ,PNEUMONIA ,HEAT shock proteins ,MOLECULAR chaperones ,LUNG diseases ,PATIENTS ,MEDICAL research - Abstract
Background: Heat shock protein (HSP) 47, a collagen-specific molecular chaperone, is involved in the processing and/or secretion of procollagens, and its expression is increased in various fibrotic diseases. The aim of this study was to determine whether quantitative immunohistochemical evaluation of the expression levels of HSP47, type I procollagen and α-smooth muscle actin (SMA) allows the differentiation of idiopathic usual interstitial pneumonia (UIP) from UIP associated with collagen vascular disease (CVD) and idiopathic nonspecific interstitial pneumonia (NSIP). Methods: We reviewed surgical lung biopsy specimens of 19 patients with idiopathic UIP, 7 with CVD-associated UIP and 16 with idiopathic NSIP and assigned a score for the expression of HSP47, type I procollagen and α-SMA in type II pneumocytes and/or lung fibroblasts (score 0 = no; 1 = weak; 2 = moderate; 3 = strong staining). Results: The expression level of HSP47 in type II pneumocytes of idiopathic UIP was significantly higher than in CVD-associated UIP and idiopathic NSIP. The expression of HSP47 in fibroblasts was significantly higher in idiopathic UIP and idiopathic NSIP than in CVD-associated UIP. The expression of type I procollagen in type II pneumocytes was significantly higher in idiopathic UIP than in idiopathic NSIP. The expression of type I procollagen in fibroblasts was not different in the three groups, while the expression of a-SMA in fibroblasts was significantly higher in idiopathic UIP than in idiopathic NSIP. Conclusion: Our results suggest the existence of different fibrotic pathways among these groups involved in the expression of HSP47 and type I procollagen. [ABSTRACT FROM AUTHOR]
- Published
- 2005
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16. Crystallization and preliminary crystallographic studies of the butyrolactone autoregulator receptor protein (BarA) from Streptomyces virginiae.
- Author
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Yoon, Young-Ho, Kawai, Fumihiro, Sugiyama, Kanako, Park, Sam-Yong, Nihira, Takuya, Choi, Sun-Uk, and Hwang, Yong-Il
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BACTERIAL protein crystallography ,BUTYROLACTONES ,STREPTOMYCES ,CRYSTALLIZATION ,DNA-binding proteins ,VIRGINIAMYCIN - Abstract
The Streptomyces butyrolactone autoregulator receptor protein (BarA) is a DNA-binding protein that regulates the biosynthesis of the antibiotic virginiamycin. In this study, BarA from S. virginiae was overexpressed in Escherichia coli, purified and crystallized. Crystals of purified protein have been grown that diffracted to beyond 3.0 Å resolution at 100 K using synchrotron radiation. The protein crystals belonged to the hexagonal space group P6
5 22, with unit-cell parameters a = b = 128.0, c = 286.2 Å. With four molecules per asymmetric unit, the crystal volume per unit protein mass ( VM ) was 3.2 Å3 Da−1 and the solvent content was 62%. [ABSTRACT FROM AUTHOR]- Published
- 2010
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17. Structural characterization of the photoswitchable fluorescent protein Dronpa-C62S
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Nam, Ki-Hyun, Kwon, Oh Yeun, Sugiyama, Kanako, Lee, Won-Ho, Kim, Young Kwan, Song, Hyun Kyu, Kim, Eunice Eunkyung, Park, Sam-Yong, Jeon, Hyesung, and Hwang, Kwang Yeon
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GREEN fluorescent protein , *FLUORESCENT polymers , *GENETIC mutation , *HYDROGEN bonding - Abstract
Abstract: The photoswitching behavior of green fluorescent proteins (GFPs) or GFP-like proteins is increasingly recognized as a new technique for optical marking. Recently, Ando and his colleagues developed a new green fluorescent protein Dronpa, which possesses the unique photochromic property of being photoswitchable in a non-destructive manner. To better understand this mechanism, we determined the crystal structures of a new GFP Dronpa and its mutant C62S, at 1.9Å and 1.8Å, respectively. Determination of the structures demonstrates that a unique hydrogen-bonding network and the sulfur atom of the chromophore are critical to the photoswitching property of Dronpa. Reversible photoswitching was lost in cells expressing the Dronpa-C62S upon repetitive irradiation compared to the native protein. Structural and mutational analyses reveal the chemical basis for the functional properties of photoswitchable fluorescent proteins and provide the basis for subsequent coherent engineering of this subfamily of Dronpa homolog’s. [Copyright &y& Elsevier]
- Published
- 2007
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18. G196 epitope tag system: a novel monoclonal antibody, G196, recognizes the small, soluble peptide DLVPR with high affinity.
- Author
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Tatsumi K, Sakashita G, Nariai Y, Okazaki K, Kato H, Obayashi E, Yoshida H, Sugiyama K, Park SY, Sekine J, and Urano T
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- Amino Acid Sequence, Animals, Antibodies, Monoclonal biosynthesis, Antibodies, Monoclonal isolation & purification, Antibody Affinity, Antibody Specificity, Binding Sites, Cloning, Molecular, Crystallography, X-Ray, Epitopes genetics, Epitopes immunology, Escherichia coli genetics, Escherichia coli metabolism, Gene Expression, Genetic Vectors chemistry, Genetic Vectors metabolism, HeLa Cells, Humans, Mice, Peptides genetics, Peptides immunology, Protein Binding, Protein Conformation, alpha-Helical, Protein Conformation, beta-Strand, Protein Interaction Domains and Motifs, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins immunology, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, Antibodies, Monoclonal chemistry, Epitope Mapping methods, Epitopes chemistry, Peptides chemistry
- Abstract
The recognition specificity of monoclonal antibodies (mAbs) has made mAbs among the most frequently used tools in both basic science research and in clinical diagnosis and therapies. Precise determination of the epitope allows the development of epitope tag systems to be used with recombinant proteins for various purposes. Here we describe a new family of tag derived from the epitope recognized by a highly specific mAb G196. The minimal epitope was identified as the five amino acid sequence Asp-Leu-Val-Pro-Arg. Permutation analysis was used to characterize the binding requirements of mAb G196, and the variable regions of the mAb G196 were identified and structurally analyzed by X-ray crystallography. Isothermal titration calorimetry revealed the high affinity (K
d = 1.25 nM) of the mAb G196/G196-epitope peptide interaction, and G196-tag was used to detect several recombinant cytosolic and nuclear proteins in human and yeast cells. mAb G196 is valuable for developing a new peptide tagging system for cell biology and biochemistry research.- Published
- 2017
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19. The crystal structure of the active domain of Anopheles anti-platelet protein, a powerful anti-coagulant, in complex with an antibody.
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Sugiyama K, Iyori M, Sawaguchi A, Akashi S, Tame JR, Park SY, and Yoshida S
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- Amino Acid Sequence, Animals, Anticoagulants chemistry, Anticoagulants immunology, Base Sequence, Blood Coagulation, Cloning, Molecular, Crystallography, X-Ray, DNA Primers, Models, Molecular, Molecular Sequence Data, Mutagenesis, Polymerase Chain Reaction, Protein Conformation, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Anopheles metabolism, Antibodies immunology, Anticoagulants metabolism
- Abstract
Blood clotting is a vitally important process that must be carefully regulated to prevent blood loss on one hand and thrombosis on the other. Severe injury and hemophilia may be treated with pro-coagulants, whereas risk of obstructive clotting or embolism may be reduced with anti-coagulants. Anti-coagulants are an extremely important class of drug, one of the most widely used types of medication, but there remains a pressing need for novel treatments, however, as present drugs such as warfarin have significant drawbacks. Nature provides a number of examples of anti-coagulant proteins produced by blood-sucking animals, which may provide templates for the development of new small molecules with similar physiological effects. We have, therefore, studied an Anopheles anti-platelet protein from a malaria vector mosquito and report its crystal structure in complex with an antibody. Overall the protein is extremely sensitive to proteolysis, but the crystal structure reveals a stable domain built from two helices and a turn, which corresponds to the functional region. The antibody raised against Anopheles anti-platelet protein prevents it from binding collagen. Our work, therefore, opens new avenues to the development of both novel small molecule anti-clotting agents and anti-malarials., (© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2014
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20. Elevated levels of interferon gamma-inducible protein-10 and epithelial neutrophil-activating peptide-78 in patients with pulmonary sarcoidosis.
- Author
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Sugiyama K, Mukae H, Ishii H, Kakugawa T, Ishimoto H, Nakayama S, Shirai R, Fujii T, Mizuta Y, and Kohno S
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- Adult, Aged, CD4-Positive T-Lymphocytes pathology, CD8-Positive T-Lymphocytes pathology, Case-Control Studies, Cell Count, Cell Movement, Chemokine CXCL10, Chemokine CXCL5, Chemokines, CXC analysis, Chemokines, CXC genetics, Female, Gene Expression Regulation, Humans, Male, Middle Aged, Receptors, Interleukin-2 analysis, Receptors, Interleukin-2 genetics, Receptors, Interleukin-2 metabolism, Sarcoidosis, Pulmonary pathology, Bronchoalveolar Lavage Fluid chemistry, Bronchoalveolar Lavage Fluid cytology, Chemokines, CXC metabolism, Sarcoidosis, Pulmonary metabolism
- Abstract
Objective and Background: Interferon gamma-inducible protein (IP)-10 and epithelial neutrophil-activating peptide (ENA)-78 belong to the CXC chemokine family and are important factors in inflammatory lung diseases. In sarcoidosis, the potential role of IP-10 to regulate the migration and activation of T-cells towards sites of sarcoid activity has been suggested., Methods: In this study, the concentrations of IP-10 and ENA-78 in the serum and BAL fluid of patients with different stages of active pulmonary sarcoidosis (n=41) and healthy subjects (n=12) were measured by enzyme-linked immunosorbent assay to evaluate the contribution of these CXC chemokines to this disease., Results: Serum and BAL fluid concentrations of IP-10 and BAL fluid levels of ENA-78 in patients with sarcoidosis were significantly higher than those in control subjects. The serum levels of IP-10 were significantly increased only in patients with stages I and II sarcoidosis, while BAL fluid levels of ENA-78 were increased only in stage III sarcoidosis. In addition, serum concentrations of IP-10 were elevated in patients with extrapulmonary lesions compared with those without such lesions. In patients with sarcoidosis, IP-10 concentrations in BAL fluid correlated with lymphocyte proportions in BAL fluid., Conclusion: IP-10 may play an important role in regulating lymphocytes into the lung and that ENA-78 may be associated with lung parenchymal disease in pulmonary sarcoidosis.
- Published
- 2006
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21. Differential effects of alpha- and beta-defensin on cytokine production by cultured human bronchial epithelial cells.
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Sakamoto N, Mukae H, Fujii T, Ishii H, Yoshioka S, Kakugawa T, Sugiyama K, Mizuta Y, Kadota J, Nakazato M, and Kohno S
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- Bronchi cytology, Cell Survival drug effects, Cells, Cultured, Cytokines genetics, Epithelial Cells drug effects, Epithelial Cells metabolism, Humans, NF-kappa B metabolism, RNA, Messenger metabolism, Bronchi drug effects, Bronchi metabolism, Cytokines biosynthesis, alpha-Defensins pharmacology, beta-Defensins pharmacology
- Abstract
Defensins are cysteine-rich cationic antimicrobial peptides that play an important role in innate immunity and are known to contribute to the regulation of host adaptive immunity. In addition to direct antimicrobial activities, it has been recently reported that alpha-defensins, mainly present in neutrophils in the lung, have a cytotoxic effect and induce IL-8 production from airway epithelial cells. Although beta-defensins are expressed in epithelial cells in various tissues, including lung, there are no reports of their effects on cytokine synthesis in airway epithelial cells. The aim of the present study was to determine the effects of both alpha- and beta-defensins on the cytokine production, transcription factor binding activity, and cytotoxicity in primary cultured human bronchial epithelial cells (HBECs). We used human neutrophil peptide-1 (HNP-1; alpha-defensin) and human beta-defensin-2 (HBD-2) to stimulate HBECs. The results showed that treatment of HBECs with HNP-1, but not HBD-2, increased IL-8 and IL-1beta mRNA expression in a dose-dependent manner and also enhanced IL-8 protein secretion and NF-kappaB DNA binding activity. The 24-h treatments with >20 microg/ml of HNP-1 or >50 microg/ml of HBD-2 were cytotoxic to HBECs. These results suggest that alpha- and beta-defensins have different effects on cytokine synthesis by airway epithelial cells, and we speculate that they play different roles in inflammatory lung diseases.
- Published
- 2005
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22. High-BAL fluid concentrations of RANTES in nonspecific interstitial pneumonia compared with usual interstitial pneumonia.
- Author
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Yoshioka S, Mukae H, Sugiyama K, Kakugawa T, Sakamoto N, Nakayama S, Abe K, Fujii T, Kadota J, and Kohno S
- Subjects
- Bronchoalveolar Lavage Fluid cytology, Cell Count, Chemokine CCL4, Female, Humans, Male, Middle Aged, Bronchoalveolar Lavage Fluid chemistry, Chemokine CCL2 analysis, Chemokine CCL5 analysis, Lung Diseases, Interstitial metabolism, Macrophage Inflammatory Proteins analysis
- Abstract
Chemokines such as regulated on activation, normal T-cell expressed and secreted (RANTES), monocyte chemoattractant protein (MCP)-1, monocyte inflammatory protein (MIP)-lalpha have been reported to play an important role in the pathogenesis of interstitial lung diseases. Among idiopathic interstitial pneumonia (IIP), nonspecific interstitial pneumonia (NSIP) has elevated percentages of Lymphocytes in bronchoalveolar lavage (BAL) fluid compared with usual interstitial pneumonia (UIP). These chemokines are candidate mediators for lymphocyte attraction to the lung in NSIR Therefore, we measured the BAL fluid levels of RANTES, MCP-1 and MIP1-alpha in 15 patients with idiopathic NSIP, 20 with idiopathic UIP, 22 with sarcoidosis and 12 healthy volunteers to evaluate the contribution of these chemokines using enzyme-linked immunosorbent assays. The levels of RANTES in BAL fluid were significantly higher in patients with NSIP compared with healthy volunteers (P < 0.01), UIP and sarcoidosis (P < 0.05). In MCP-1, the levels in BAL fluid of NSIP and UIP patients were significantly elevated compared with healthy volunteers and sarcoidosis patients (P < 0.01). These results suggest that RANTES and MCP-1 in BAL fluid may play an important role in inflammatory cell recruitment to the lung in idiopathic NSIP as well as other interstitial lung diseases.
- Published
- 2004
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