Production of monoclonal antibodies against interleukin-1α and -1β

Grassi, Jacques; Frobert, Yveline; Pradelles, Philippe; Chercuitte, Francine; Gruaz, Dominique; Dayer, Jean‐Michel; Poubelle, Patrice E.

doi:10.1016/0022-1759(89)90223-8

Cited by 135 publications

(42 citation statements)

References 29 publications

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“…Analyzis of the signal peptides [3, 4, 341 shows that their lengths and general features are similar to those of stroma-directed polypeptides such as ferredoxin or the small subunit of ribulose-bisphosphate carboxylase. In addition, the corresponding polypeptides in the cyanobacterium Synechocystis 6803 are synthesized without a signal peptide, indicating that they do not cross the thylakoid membrane [9, 351. Second, PSI polypeptides of 19 …”

Section: Discussionmentioning

confidence: 99%

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Purification and membrane topology of PSI‐D and PSI‐E, two subunits of the photosystem I reaction center

Lagoutte

Valloni

1992

European Journal of Biochemistry

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Structural studies have been conducted on polypeptides PSI-D and PSI-E, whch are extrinsic but firmly bound to the photosystem I reaction center. These subunits are predicted to be involved in the correct interaction with soluble electron acceptor(s), like ferredoxin. We designed an original method to extract both polypeptides directly from thylakoid membranes and to purify them: a stepwise extraction with NaSCN followed by size fractionation and reverse-phase HPLC. Investigation of the in situ topology of PSI-D and PSI-E was undertaken using monoclonal antibody binding, controlled proteolysis, peptide sequencing and electron microscopy. The precise identification of numerous proteolytic sites indicates that the entire N-terminal regions of PSI-E (up to Glu15) and PSI-D (up to LyslS) are exposed to the medium. Partial mapping of the exposed epitopes was possible using purified fragments of each polypeptide. In the case of PSI-E, this mapping confirmed the accessibility of the N-terminal part, and suggested the need for another exposed sequence, probably located after Met39 in the second half of the protein. For PSI-D, this mapping revealed that the sequence between Met74 and Metl40, including the most basic amino acid clusters, is also partly accessible. These experiments provide the first detailed informations, although still partial, on the topology of these polypeptides. They give a preliminary basis for hypotheses concerning the sites of interaction with the soluble counterparts.

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Section: Discussionmentioning

confidence: 99%

“…Tests for complementarity [19] of antibodies required the purification of the immunoglobulin fraction. This was done by double-differential precipitation with octanoic acid and ammonium sulfate (201.…”

Section: Solid-phase Immunological Testsmentioning

confidence: 99%

Purification and membrane topology of PSI‐D and PSI‐E, two subunits of the photosystem I reaction center

Lagoutte

Valloni

1992

European Journal of Biochemistry

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“…Mice were immunized with the purified recombinant protein RECA.His. Immunization of mice, cell fusion, and screening of the hybridoma supernatants by a specific enzyme immunoassay using RECA.His either as immunogen or biotin-labeled as tracer were performed as described previously (31). The positive supernatants were analyzed further by immunocytochemistry on HEK 293 cells expressing human ALK protein.…”

Section: Methodsmentioning

confidence: 99%

Activation and Inhibition of Anaplastic Lymphoma Kinase Receptor Tyrosine Kinase by Monoclonal Antibodies and Absence of Agonist Activity of Pleiotrophin

Moog-Lutz

Degoutin

Gouzi

et al. 2005

Journal of Biological Chemistry

145

168

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Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase that is transiently expressed in specific regions of the central and peripheral nervous systems, suggesting a role in its normal development and function. The nature of the cognate ligands of ALK in vertebrate is still a matter of debate. We produced a panel of monoclonal antibodies (mAbs) directed against the extracellular domain of the human receptor. Two major species of ALK (220 and 140 kDa) were identified in transfected cells, and the use of our mAbs established that the 140-kDa species results from a cleavage of the 220-kDa form. Two mAbs, in the nM range, induced the differentiation of PC12 cells transiently transfected with ALK. In human embryonic kidney 293 cells stably expressing ALK, these two mAbs strongly activated the receptor and subsequently the mitogen-activated protein kinase pathway. We further showed for the first time that activation of ALK also resulted in a specific activation of STAT3. In contrast, other mAbs presented the characteristics of blocking antibodies. Finally, in these cell systems, a mitogenic form of pleiotrophin, a proposed ligand of ALK, failed to activate this receptor. Thus, in the absence of clearly established ligand(s) in vertebrates, the availability of mAbs allowing the activation or the inhibition of the receptor will be essential for a better understanding of the biological roles of ALK. Receptors tyrosine kinase (RTKs)1 play essential roles during the development of the nervous system, regulating a wide range of cellular processes such as proliferation, survival, differentiation, and synaptogenesis. Generally, after ligand binding, RTK dimerizes, autophosphorylates, and initiates signal transduction cascades that subsequently lead to cellular responses (for review, see Ref. 1).Anaplastic lymphoma kinase (ALK) was originally identified as a RTK that acquires transforming capability when truncated and fused in the t(2;5) chromosomal rearrangement associated with the non-Hodgkin lymphoma (2). This translocation produces a fusion gene that encodes a soluble chimeric transforming protein comprising the N-terminal portion of the phosphoprotein nucleophosmin (NPM) linked to the cytoplasmic portion of ALK. It has been demonstrated that the NPM portion is responsible for the dimerization of the fusion protein leading to the constitutive activation of the kinase and to the transforming activity. Phospholipase C␥, phosphatidylinositol 3-kinase, STATs, and Src appear to be important downstream targets of NPM-ALK which contribute to its mitogenic and antiapoptotic activities (3-7). ALK is also involved in different variant chromosomal translocations (for review, see Ref. 8), all leading to the expression of fusion proteins exhibiting a constitutive activation of the kinase.Human, mouse, and Drosophila cDNAs encoding full-length ALK have been characterized (9 -11). The deduced amino acid sequences revealed that ALK is a novel RTK having an extracellular domain, a single transmembrane domain, and an intracellular do...

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“…Purified immunoglobulin from rabbit polyclonal antibody anti-His 6 -HSA kin17 (IgG 77P) was obtained as described elsewhere. 4 A conventional two-site immunometric assay (sandwich immunoassay) based on two specific monoclonal antibodies recognizing nonoverlapping epitopes was developed essentially as described by Grassi et al (24). We chose the mAb K3 and mAb K31 monoclonal antibodies, the FIG.…”

Section: Methodsmentioning

confidence: 99%

“…To quantify the amount of HSA kin17 protein/cell, we used a two-site immunometric assay (sandwich immunoassay) based on two monoclonal antibodies (24). HSA kin17 recombinant protein allowed us to calibrate the ELISA (data not shown).…”

Section: Fig 5-continuedmentioning

confidence: 99%

Ionizing Radiation Triggers Chromatin-bound kin17 Complex Formation in Human Cells

Biard¹,

Miccoli²,

Despras³

et al. 2002

Journal of Biological Chemistry

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The human DNA-binding HSA kin17 protein cross-reacts with antibodies raised against the stress-activated Escherichia coli RecA protein. We show here that HSA kin17 protein is directly associated with chromosomal DNA as judged by cross-linking experiments on living cells. We detected increased amounts of DNAbound HSA kin17 protein 24 h after ␥ irradiation, with 2.6-fold more HSA kin17 molecules after 6 Gy of irradiation (46,000 -117,000 molecules). At this time we observed that highly proliferating RKO cells displayed the concentration and co-localization of HSA kin17 and replication protein A in nucleoplasmic foci. Our results suggest that 24 h post-irradiation HSA kin17 protein may localize at the sites of unrepaired DNA damages. RKO clones expressing an HSA KIN17 antisense transcript (RASK.5 and RASK.13 cells) revealed that reduced HSA kin17 protein levels are correlated with a decrease in clonogenic cell growth and cell proliferation, as well as an accumulation of cells in early and mid-S phase. Taken together our observations support the idea that HSA kin17 protein is a DNA maintenance protein involved in the cellular response to the presence of DNA damage and suggest that it helps to overcome the perturbation of DNA replication produced by unrepaired lesions. Ionizing radiation (IR)1 induces a large range of DNA damage, including DNA double-strand breaks (DSBs), which represent a major threat to the integrity of mammalian genomes through chromosomal breakages and rearrangements (1). In mammalian cells, DSBs are repaired either by the homologous recombinational repair or by nonhomologous end joining (2, 3). DSB repair pathways are usually characterized by the sequestration of many factors into discrete nuclear foci at the sites of DNA lesions and until completion of DSB repair (4). Some of the proteins belonging to these pathways act as a sensor for DNA damage or are involved in cell cycle checkpoints. This is the case for the histone H2AX, 53BP1, RPA, Rad51, BRCA1, or the Mre11-Rad50-Nbs1 nuclease complex (4 -8). For instance, the tumor suppressor gene BRCA1, previously involved in the regulation of the replication checkpoint and transcription-coupled repair, forms a multiprotein complex with Mre11-Rad50-Nbs1 and other proteins following irradiation, termed as BASC (BRCA1-associated surveillance complex), which may serve as a sensor of DNA lesions (8,9).In this cascade of IR-induced proteins forming nuclear foci, we characterized here the HSA kin17 protein. Murine MMU kin17 protein was identified on the basis of a cross-reactivity with antibodies raised against the Escherichia coli RecA protein, a key enzyme in homologous recombination and recombinational repair of damaged DNA (10, 11). This cross-reactivity stemmed from a sequence homology stretching over 39 amino acids highly conserved during evolution (12). This domain is located in the carboxyl-terminal region of the E. coli RecA protein, a region involved in the regulation of DNA binding (13). Recent data show that kin17 proteins are highly conserved d...

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Production of monoclonal antibodies against interleukin-1α and -1β

Cited by 135 publications

References 29 publications

Purification and membrane topology of PSI‐D and PSI‐E, two subunits of the photosystem I reaction center

Purification and membrane topology of PSI‐D and PSI‐E, two subunits of the photosystem I reaction center

Activation and Inhibition of Anaplastic Lymphoma Kinase Receptor Tyrosine Kinase by Monoclonal Antibodies and Absence of Agonist Activity of Pleiotrophin

Ionizing Radiation Triggers Chromatin-bound kin17 Complex Formation in Human Cells

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