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Urinary-type plasminogen activator (uPA) expression and uPA receptor localization are regulated by α₃β₁ integrin in oral keratinocytes

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第31期275卷 23869-23876页

作者： Ghosh, S Brown, R Jones, JCR Ellerbroek, SM Stack, MS Northwestern Univ Sch Med Dept Obstet & Gynecol Chicago IL 60611 USA Northwestern Univ Sch Med Dept Cell & Mol Biol Chicago IL 60611 USA Northwestern Univ Sch Med Robert H Lurie Comprehens Canc Ctr Chicago IL 60611 USA

Expression of urinary-type plasminogen activator (uPA) and its receptor (uPAR) is correlated with matrix proteolysis, cell adhesion, motility, and invasion. To evaluate the functional link between adhesion and proteolysis in gingival keratinocytes (pp126), cells were treated with immobilized integrin antibodies to induce integrin clustering Clustering of alpha(3) and beta(1) integrin subunits, but not alpha(2), alpha(5), alpha(6), or beta(4), enhanced uPA secretion. Bead-immobilized laminin-5 and collagen I, two major alpha(3)beta(1) ligands, also induced uPA expression. Coordinate regulation of the serpin plasminogen activator inhibitor 1 was also apparent;however, a net increase in uPA activity was predominant. alpha(3)beta(1) integrin clustering induced extracellular signal-regulated kinase 1/2 phosphorylation, and both uPA induction and extracellular signal-regulated kinase activation were blocked by the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase inhibitor PD98059, Integrin aggregation also promoted a dramatic redistribution of uPAR on the cell surface to sites of clustered alpha(3)beta(1) integrins. Co-immunoprecipitation of beta(1) integrin with uPAR provided further evidence that protein-protein interactions between uPAR and beta(1) integrin control uPAR distribution, As a functional consequence of uPA up-regulation and uPA-mediated plasminogen activation, the globular domain of the laminin-5 alpha(3) subunit, a major pp126 matrix protein, was proteolytically processed from a 190-kDa form to a 160-kDa species, Laminin-5 containing the 160-kDa alpha(3) subunit efficiently nucleates hemidesmosome formation and reduces cell motility, Together, these data suggest that multivalent aggregation of the alpha(3)beta(1) integrin regulates proteinase expression, matrix proteolysis, and subsequent cellular behavior.

关键词：钙-钙调素依赖性蛋白激酶类/拮抗剂和抑制剂细胞黏附分子/代谢细胞区室化酶抑制剂/药理学类黄酮物质/药理学基因表达调控肿瘤牙龈/细胞学牙龈/代谢整合素α3β1 整合素类/代谢细胞间接合部/代谢角蛋白细胞/代谢丝裂原活化蛋白激酶1/拮抗剂和抑制剂丝裂原活化蛋白激酶1/代谢丝裂原活化蛋白激酶3 丝裂原激活蛋白激酶类/拮抗剂和抑制剂丝裂原激活蛋白激酶类/代谢磷酰化纤维蛋白溶酶原/代谢纤溶酶原激活物抑制物1 癌前状态受体细胞表面/分离和提纯肿瘤细胞培养的人类

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ISOLATION, PURIFICATION, AND BIOCHEMICAL-CHARACTERIZATION OF CALCITONIN GENE RELATED PEPTIDE RECEPTORS

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ANNALS OF THE NEW YORK ACADEMY OF SCIENCES-SERIES 1992年第1期657卷 70-87页

作者： WIMALAWANSA, SJ Biochemical Endocrinology Unit Department of Medicine and Chemical Pathology Royal Postgraduate Medical School London W12 ONN England

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Identification of a receptor for Reg (Regenerating gene) protein, a pancreatic β-cell regeneration factor

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第15期275卷 10723-10726页

作者： Kobayashi, S Akiyama, T Nata, K Abe, M Tajima, M Shervani, NJ Unno, M Matsuno, S Sasaki, H Takasawa, S Okamoto, H Tohoku Univ Grad Sch Med Dept Biochem Aoba Ku Sendai Miyagi 9808575 Japan Tohoku Univ Grad Sch Med Dept Surg Aoba Ku Sendai Miyagi 9808575 Japan Tohoku Univ Grad Sch Med Dept Geriatr Med Aoba Ku Sendai Miyagi 9808575 Japan

Reg (regenerating gene) was isolated as a gene specifically expressed in regenerating islets (Terazono, K., Yamamoto, H., Takasawa, S., Shiga, K., Yonemura, Y., Tochino, Y., and Okamoto, H. (1988) J. Biol. Chem, 263, 2111-2114), Rat and human Reg gene products, Reg/REG proteins, have been demonstrated to stimulate islet beta-cell growth in vitro and in vivo and to ameliorate experimental diabetes. In the present study, we isolated a cDNA for the Reg protein receptor from a rat islet cDNA library. The cDNA encoded a cell surface 919-amino acid protein, and the cells into which the cDNA had been introduced bound Reg protein with high affinity. When the cDNA was introduced into RINm5F cells, a pancreatic beta-cell line that shows Reg-dependent growth, the transformants exhibited significant increases in the incorporation of 5'-bromo-2'-deoxyuridine as well as in the cell numbers in response to Reg protein. A homology search revealed that the cDNA is a homologue to a human multiple exostoses-like gene, the function of which has hitherto been unknown. These results strongly suggest that the receptor is encoded by the exostoses-like gene and mediates a growth signal of Reg protein for beta-cell regeneration.

关键词：氨基酸序列 CHO细胞 COS细胞钙结合蛋白质类/代谢仓鼠亚科 DNA 互补/分离和提纯胰岛/代谢胰石蛋白分子序列数据神经组织蛋白质类受体细胞表面/分离和提纯再生动物人类大鼠

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Differences in the polar clustering of the high- and low-abundance chemoreceptors of Escherichia coli

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 2000年第14期97卷 8057-8062页

作者： Lybarger, SR Maddock, JR Univ Michigan Dept Biol Ann Arbor MI 48109 USA

The chemosensory complexes in Escherichia coli are localized predominantly in large aggregates at one or both of the cell poles, however, neither the role of the polar localization nor the role of the clustering is understood. In E. coli, the two classes of chemoreceptors or transducers, high- and low-abundance, differ in their ability to support chemotaxis when expressed as the sole chemoreceptor type in the cell. In this study, we examined both the contribution of individual chemoreceptors to polar clustering and the ability of each chemoreceptor type to cluster in the absence of all others. We found that polar clustering of methyl-accepting chemotaxis proteins (MCPs) is not dependent on any one chemoreceptor type. Remarkably, when expressed individually at similar levels, the chemoreceptors display differential clustering abilities. The high-abundance transducers cluster at the cell pole almost as well as do the MCPs in cells expressing all four species, whereas the low-abundance transducers, although polar, are not particularly clustered. CheA and chew distributions in strains expressing only one chemoreceptor type coincide with MCP localization, indicating that the low-abundance chemoreceptors are competent for ternary complex formation but are defective in aggregation. These studies reveal that, in contrast to our previous model, polarity of the chemoreceptors is independent of clustering, suggesting that the polar localization of the chemoreceptors is not simply caused by diffusion limitations on large protein aggregates.

关键词：细菌蛋白质类细胞区室化细胞极性大肠杆菌/超微结构膜蛋白质类/分离和提纯显微镜检查免疫电子受体细胞表面/分离和提纯信号传导

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COMPARISON OF A CALCITONIN GENE RELATED PEPTIDE RECEPTOR IN A HUMAN NEUROBLASTOMA CELL-LINE (SK-N-MC) AND A CALCITONIN RECEPTOR IN A HUMAN BREAST-CARCINOMA CELL-LINE (T47D)

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ANNALS OF THE NEW YORK ACADEMY OF SCIENCES-SERIES 1992年第1期657卷 106-116页

作者： MUFF, R STANGL, D BORN, W FISCHER, JA UNIV ZURICH DEPT MED CH-8008 ZURICH SWITZERLAND

A specific CGRP-binding protein of M(r) 60,000 has been identified in the human neuroblastoma cell line SK-N-MC. After N-deglycosylation a M(r) of 48,000 was found. The M(r) were indistinguishable from those determined in the human cerebellum. Receptor binding of CGRP is coupled to cyclic AMP formation. The latter is antagonized by hCGRP-I8-37. CT and DAPamide interact only minimally with the CGRP receptor, whereas CGRP and DAPamide are full agonists in T47D cells. The CT receptor on human breast cancer cell line T47D is clearly different from the human CGRP receptor.

关键词：氨基酸序列乳腺肿瘤降钙素基因相关肽/代谢细胞系环AMP/代谢动力学分子序列数据分子量神经母细胞瘤受体降钙素受体细胞表面/遗传学受体细胞表面/分离和提纯受体细胞表面/代谢序列同源性核酸女(雌)性人类

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Molecular basis of lutropin recognition by the mannose/GalNAc-4-SO₄ receptor

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 2000年第18期97卷 9949-9954页

作者： Roseman, DS Baenziger, JU Washington Univ Sch Med Dept Pathol St Louis MO 63110 USA

The circulatory half-life of the glycoprotein hormone lutropin (LH) is precisely regulated by the mannose (Man)/GalNAc-4-SO4 receptor expressed in hepatic endothelial cells. Rapid clearance from the circulation contributes to the episodic rise and fall of LH levels that is essential for maximal stimulation of the G protein-coupled LH receptor. We have defined two molecular forms of the Man/GalNAc-4-SO4 receptor that differ in ligand specificity, cell and tissue expression, and function. The form expressed by hepatic endothelial cells binds GalNAc-4-SO4-bearing ligands and regulates hormone circulatory half-life, whereas the form expressed by macrophages binds Man-bearing ligands and may play a role in innate immunity. We demonstrate that the GalNAc-4-SO4-specific form in hepatic endothelial cells is dimeric whereas the Man-specific form in lung macrophages is monomeric, accounting for the different ligand specificities of the receptor expressed in these tissues. Two cysteine-rich domains, each of which binds a single GalNAc-4-SO4, are required to form stable complexes with LH. The kinetics of LH binding by the GalNAc-4-SO4-specific form of the receptor in conjunction with its rate of internalization from the cell surface make it likely that only two of the four terminal GalNAc-4-SO4 moieties present on native LH are engaged before receptor internalization. As a result, the rate of hormone clearance will remain constant over a wide range of LH concentrations and will not be sensitive to variations in the number of terminal GalNAc-4-SO4 moieties as long as two or more are present on multiple oligosaccharides.

关键词： CHO细胞仓鼠亚科二聚作用内皮/代谢动力学肝/代谢肺/代谢促黄体激素/化学促黄体激素/代谢甘露糖结合凝集素类模型分子蛋白质构象受体细胞表面/化学受体细胞表面/分离和提纯受体细胞表面/代谢受体 LH/代谢重组融合蛋白质类/化学重组融合蛋白质类/代谢表面等离激元转染动物大鼠

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CHARACTERIZATION OF CGRP(1) AND CGRP(2) RECEPTOR SUBTYPES

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ANNALS OF THE NEW YORK ACADEMY OF SCIENCES-SERIES 1992年第1期657卷 88-105页

作者： QUIRION, R VANROSSUM, D DUMONT, Y STPIERRE, S FOURNIER, A UNIV QUEBEC INST NATL RECH SCI SANTE POINTE CLAIRE H3R 1G6 QUEBEC CANADA MCGILL UNIV DOUGLAS HOSP RES CTR DEPT PHARMACOL & THERAPEUT VERDUN H34 1R3 QUEBEC CANADA

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A NEW PATHWAY FOR VITAMIN-A - UNDERSTANDING THE PLEIOTROPIC EFFECTS OF RETINOIDS

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ANNALS OF THE NEW YORK ACADEMY OF SCIENCES 1992年第1期669卷 70-86页

作者： LEVIN, AA STURZENBECKER, LJ KAZMER, S BOSAKOWSKI, T HUSELTON, C ALLENBY, G SPECK, J KRATZEISEN, C ROSENBERGER, M LOVEY, A GRIPPO, JF HOFFMANN LA ROCHE INC DEPT MED CHEM NUTLEY NJ 07110 USA HOFFMANN LA ROCHE INC DEPT DRUG METAB NUTLEY NJ 07110 USA

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MULTIPLE FOLATE TRANSPORT-SYSTEMS IN L1210 CELLS

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ADVANCES IN ENZYME REGULATION 1992年第C期32卷 3-+页

作者： FAN, J VITOLS, KS HUENNEKENS, FM DepartmentofMolecularandExperimentalMedicine ScrippsResearchInstituteLaJollaCA92037.

Biotin derivatives of methotrexate (biotin-SS-MTX) and folate (biotin-SS-folate), in which the functional components are joined by a dissociable disulfide-containing spacer, have been synthesized, purified by DEAE-Trisacryl chromatography, and characterized by HPLC, elemental analysis and mass spectrometry. These compounds provide a convenient means for the single-step purification of the folate transporters from L1210 cells. Parental L1210 murine leukemia cells, which contain only the microM transporter (the reduced folate/MTX transport protein) were treated with the N-hydroxysulfosuccinimide ester of biotin-SS-MTX, and a detergent extract of the plasma membranes was exposed to streptavidin-agarose beads to adsorb the labeled protein. Dithiothreitol cleavage of the disulfide linkage released the transporter, which migrated as a well-defined component (43 kDa) on SDS-PAGE gels; no other proteins were present. An L1210 subline (JF), obtained by adapting cells to grow on nanomolar concentrations of folate, contains both the microM transporter and the nM transporter (high-affinity folate binding protein). When these cells were treated with the N-hydroxysulfosuccimide ester of biotin-SS-folate and processed as described above, analysis on SDS-PAGE gels revealed the presence of two proteins, the microM transporter (43 kDa) and the nM transporter (39 kDa). Both transporters were characterized with respect to amino acid content; blocked N-termini precluded Edman sequencing. Treatment of the nM transporter with peptide:N-glycosidase F produced a smaller component (32 kDa); the microM transporter, conversely, was unchanged by this procedure. When the microM transporter in parental L1210 cells was labeled with fluorescein-MTX and then treated with phosphoinositol-specific phospholipase C (PI-PLC), no change in fluorescence was detected. Alternatively, when the nM transporter in the JF subline was labeled with fluorescein-folate and then treated with PI-PLC, complete loss of f

关键词：生物素/类似物和衍生物生物素/化学合成生物素/代谢载体蛋白质类/化学载体蛋白质类/分离和提纯载体蛋白质类/代谢色谱法亲和/方法叶酸/类似物和衍生物叶酸/化学合成叶酸/代谢白血病L1210/代谢甲氨蝶呤/类似物和衍生物甲氨蝶呤/化学合成甲氨蝶呤/代谢受体细胞表面/分离和提纯受体细胞表面/代谢肿瘤细胞培养的动物小鼠

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BIOLOGICALLY-ACTIVE AMYLIN-BIOTIN AS A PROBE FOR ISOLATING AMYLIN CGRP RECEPTORS

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ANNALS OF THE NEW YORK ACADEMY OF SCIENCES-SERIES 1992年第1期657卷 452-454页

作者： CHANTRY, A FOOT, EA LEIGHTON, B MRC Immunochemistry Unit Department of Biochemistry University of Oxford Oxford OXI 3QU United Kingdom

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