巻号一覧
10 巻 (1985)
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10 巻, 3 号
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Kei Takahashi, Hiroyuki Naora1985 年 10 巻 3 号 p. 195-208
発行日: 1985年
公開日: 2008/04/18
ジャーナル フリーOur previous study showed that an epitope defined by a monoclonal antibody against human urokinase is located on the 33-Kdalton catalytic domain of the enzyme (Nakamura, M. et al., Cell Struct Funct., 9, 167-179, 1984). The epitope structure was further determined and chara-cterized on one-dimensional SDS-polyacrylamide slab gel maps of CNBr-cleaved polypeptide fragments as well as on their Western blots. A single homogeneous polypeptide with an approximate molecular weight of 3.4-Kdaltons was found to be antigenic. The monoclonal antibody exhibited a stronger inhibition of the enzyme activity than the polyclonal antibodies tested, and cross-reacted with a 65-Kdalton tissue-type plasminogen activator present in Detroit 562 cells. From these results and data made up with the help of a computer comparison of known sequences of urokinase and a tissue-type plasminogen activator, we concluded that the epitope is Cys-Gln-Gly-Asp-Ser-Gly-Gly-Pro-Leu-Val-Cys and containes a catalytically active residue, serine.抄録全体を表示PDF形式でダウンロード (6824K) -
Kei Takahashi, Takashi Gojobori, Hiroyuki Naora1985 年 10 巻 3 号 p. 209-218
発行日: 1985年
公開日: 2008/04/18
ジャーナル フリーTwelve amino acid sequences of kringle-forming polypeptides were compiled from the known sequences of urokinase A-chain (human), a tissue-type plasminogen activator (human), prothrombin (human and bo-vine), and plasminogen (human). Their sequence homologies with maximum match were examined by a computer program. A homology alignment and graphic matrix analyses did show that they had a great degree of homology. All the cystein residues responsible for the kringle structures of urokinase and the tissue-type plasminogen activator were confidently preserved as well as other proteins.
A phylogenetic tree was then reconstructed, and the A-and S-chain of bovine and human prothrombins were accounted for the measurement of the evolutionary time span. It was found that urokinase had a larger time span, as much as 60 million years (MY), than the tissue-type plasminogen activator. A common ancestral element of the kringle-related serine proteases was placed at around 500 MY ago, as old as the diversion of the α-and β-chains of hemoglobin. Thus, the kringle-families have undergone a substantial evolutionary divergence. Moreover, they can be subgrouped into three subfamilies : plasminogen activators, plasminogen, and prothrombin A-chains, the last being the most distantly diverged prothrombin S-chains.抄録全体を表示PDF形式でダウンロード (1133K) -
Monoclonal Antibody that Immunoreacts with a Subclass of Human Receptors for Epidermal Growth FactorM. Ali Behzadianl, Nobuyoshi Shimizu1985 年 10 巻 3 号 p. 219-232
発行日: 1985年
公開日: 2008/04/18
ジャーナル フリーSpleen cells from BALBc mice immunized with human epidermoid carcinoma A431 cells were fused with mouse myeloma P3NP cells. One of the isolated hybridoma lines, B4G7, secreted a monoclonal antibody of the IgG class which inhibited the binding of [125I]-EGF to A431 cells and human fibroblasts, but not to mouse 3T3 cells. This inhibition was partial (65-70 %) and Scatchard analysis of the EGF binding data suggested that the B4G7 antibody interacts preferentially with a low-affinity class of EGF re-ceptors. This monoclonal antibody specifically precipitated EGF receptors (Mr =170, 000 and 155, 000) of A431 cells which were directly crosslinked with [125I]-EGF. It also precipitated EGF receptors from cells whose surface pro-teins were labeled with 125I, from cells grown in the presence of [35S]-methionine or [32P]-orthophosphate, and from membrane fractions phosphorylated in vitro with [32P]-γ-ATP. Receptors subjected to EGF-induced phosphorylation, both in vivo and in vitro, were also precipitated. The B4G7 antibody blocked70 % of the EGF receptors in human fibroblasts, but did not stimulate DNA synthesis in these cells. However, in the presence of this antibody, cells showed the full mitogenic response to EGF, presumably through the unblocked re-ceptors that are likely to be of the high-affinity type.抄録全体を表示PDF形式でダウンロード (5835K) -
Hideki Gotoh, Toshifumi Takenaka, Hidenori Horie, Yukio Hiramoto1985 年 10 巻 3 号 p. 233-243
発行日: 1985年
公開日: 2008/04/18
ジャーナル フリーIntracellular organelle motion within clonal pituitary tumor cells (GH3) was observed directly with a contrast enhancement, computer-video microscope system. All particles except nuclei moved in a complex fashion. Two types of particles predominated; one large and round, the other small and elongated. We classified the movements of these particles as saltation, oscillation and slow translocation. Saltation was directional movement with velocity of the order of 1 μm/sec. Oscillation was local motion occurring within 1 μm that showed no specific direction. Its velocity was similar to that of saltation. Large particles, in particular, showed the 3rd type of movement, slow translocation. The velocity appeared to be one order slower than that of saltation.
We also examined the cells with fluorescent, dark-field and electron micros-copies. We concluded that the large round particles were lysosomes and the small elongated ones mitochondria. The microtubule depolymerizer, vinblastine and the microfilament depolymerizer, cytochalasin D, completely inhibited all the types of organelle movement. The mechanism and significance of these organelle movements are discussed.抄録全体を表示PDF形式でダウンロード (9532K) -
Toshimichi Yoshida, Atsushi Ito, Kosaku Izutsu1985 年 10 巻 3 号 p. 245-258
発行日: 1985年
公開日: 2008/04/18
ジャーナル フリーTwo anti-dynein-1 antibodies were affinity-purified from rabbit antiserum produced by immunization with dynein-1 from sea urchin sperms. One was prepared with dynein-1 as the ligand and the other with the A heavy chains of dynein-1 as the ligand. In Western blots of axonemal proteins, the former antibody reacted with the A heavy chains of dynein-1 as well as with several other smaller polypeptides, whereas the latter bound almost exclusively to the A heavy chains.
PtK 2 cells stained by indirect immunofluorescence with either of these antidyneins had identical fluorescence patterns. The interphase cell showed rather diffuse and weak fluorescence in its nucleus and perinuclear cytoplasm. Its primary cilium and its centrioles also fluoresced. During prophase and prometaphase, a more intense fluorescence was present in the asters and developing spindle. During metaphase and anaphase the half-spindles fluo-resced intensely in a fibrous pattern that corresponded to that of the spindle fibers, showing less intense fluorescence in the anaphase interzone. In telophase and early interphase, the intercellular bridge on each side of the midbody also was stained. These results are evidence that dynein-1, specifically the A heavy chains and/or a related antigen, is densely packed in the mitotic spindle of PtK 2 cells.抄録全体を表示PDF形式でダウンロード (1694K) -
Katsumi Yamashita, Frances M. Davis, Potu N. Rao, Mutsuo Sekiguchi, Ta ...1985 年 10 巻 3 号 p. 259-270
発行日: 1985年
公開日: 2008/04/18
ジャーナル フリーIn tsBN2 cells, a temperature-sensitive (ts) mutant of the BHK21 cell line, with a ts-defect in its regulatory system for chromosome condensation, antigens that react with mitotic specific mouse monoclonal antibody MPM-2 were produced when premature chromosome condensation (PCC) was induced by a temperature shift. The polypeptides of antigens recognized by MPM-2 in tsBN2 cells with PCC were identical to those of antigens in mitotic cells. These antigens appeared concomitantly with chro-mosome condensation, which suggests that these mitotic-specific antigens may be related to chromosome condensation. As the production of mitotic-specific antigens was inhibited by W-7, a specific and potent antagonist of calmodulin, calmodulin may function in the mitotic phosphorylation of nonhistone protein.抄録全体を表示PDF形式でダウンロード (1211K) -
Shusuke Hirano, Noriko Iwasaki-Mutou, Hiroaki Asou, Yoshihiro Ogawa1985 年 10 巻 3 号 p. 271-278
発行日: 1985年
公開日: 2008/04/18
ジャーナル フリーDeveloping cultures of dissociated cerebral hemispheres ob-tained from 18-day-old embryonic rats synthesized, or activated, a myelin-related enzyme, 2', 3'-cyclic nucleotide 3'-phosphohydrolase (CNPase). This increase in activity coincided with the onset of myelination. The presence of CNPase in oligodendrocytes and myelin was demonstrated using immuno-cytochemical staining techniques. Myelinated axons and myelin sheaths were clearly made visible by electron microscopy of 28-day-old cultures.抄録全体を表示PDF形式でダウンロード (831K) -
Mayumi Ono, Kazuaki Mannen, Tatsuo Shimada, Vlichihiko Kuwano, Kumato ...1985 年 10 巻 3 号 p. 279-294
発行日: 1985年
公開日: 2008/04/18
ジャーナル フリーWe compared the effects of the cationic ionophore, monensin, on the synthesis, maturation and release of vesicular stomatitis virus (VSV) in cultures of Chinese hamster ovary (CHO) cells and the monensin-resistant clone, MonR-31. Our results depended on the dose and time of the addition of monensin to the infected cells, from 1 h prior to VSV infection to 1 h after infection. VSV production was more resistant in MonR-31 than in CHO cells when the ionophore was added 1 h prior to VSV infection. Monensin added 1 h after VSV infection showed the opposite phenomenon; release of virus particles into the medium was 10-to 105-fold less in MonR-31 cells than in CHO cells, and the intracellular virus number in the resistant cells was one-third to one-fourth of that in the parental CHO cells. Syntheses of all virus-associated G, N and M proteins were inhibited in both cell lines by monensin, but especially so in the MonR-31 cells. There were no marked qualitative changes in the biochemical properties of viral glycoprotein G in virus-infected CHO and MonR-31 cells treated with monensin after virus infection. An endoglycosidase H-resistant G with a molecular weight smaller than that of normal G and attachments of palmitate or fucose on the truncated G protein appeared. Alteration of the secretion of as well as the synthesis of the enveloped virus is discussed in relation to the monensin susceptibility of the resistant MonR-31 clone.抄録全体を表示PDF形式でダウンロード (1640K) -
Hajime Dugihara, Nobuhisa Yonemitsu, Kankatsu Yun, Shinichi Miyabara1985 年 10 巻 3 号 p. 295-303
発行日: 1985年
公開日: 2008/04/18
ジャーナル フリーUnder lowered Ca2+ content and in the post-treatment state of low Ca2+, we studied the cellular structure and functioning in mouse adrenocortical tumor cells, Y-1. These cells had been maintained in Ham F12 medium containing 10 % fetal calf serum. The Ca2+ present in this complete medium was 0.39 mM.
Under a slightly lowered Ca2+ (0.29 mM) produced by EGTA, the cells had many blebs on their surfaces and specific functional activity decreased in steroidogenesis as did ACTH reactivity. In the post-treatment state of a low Ca2+, the cellular surface was covered with many short microvilli and there was greater cellular activity than in the control cells.
When the Ca2+ concentration was below 0.17 mM, the cellular structure and functioning were disturbed, and there was no recovery even at the physiological Ca2+ after the removal of EGTA.抄録全体を表示PDF形式でダウンロード (730K) -
Kiyoshi Ueda, Hiroshi Nakayasu1985 年 10 巻 3 号 p. 305-309
発行日: 1985年
公開日: 2008/04/18
ジャーナル フリーWe examined the distribution of actin in isolated nuclear matrices from mouse leukemia L5178Y cells using an anti-actin antibody and protein A-conjugated colloidal gold particles. Before immunogold staining, we partially digested the surface lamina of the nuclear matrix with trypsin (Nakayasu and Ueda, Exp. Cell Res. 143, 55-62, 1983) to allow penetration of the gold particles into the nuclear matrix. Trypsin digestion slightly modified the internal structure of the nuclear matrix, but did not affect the actin content in the nuclear matrix nor the reactivity of actin with the antibody. Many colloidal gold particles were present along fibrogranular structures in the nuclear matrix. The results reported here confirm the existence of actin in the interior of the nuclear matrices of L5178Y cells.抄録全体を表示PDF形式でダウンロード (515K)
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