scholarly journals Formation of the Ca2+-activated photoprotein obelin from apo-obelin and mRNA inside human neutrophils

1988 ◽  
Vol 252 (1) ◽  
pp. 143-149 ◽  
Author(s):  
A K Campbell ◽  
A K Patel ◽  
Z S Razavi ◽  
F McCapra
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Cell Activation ◽  
Membrane Attack Complex ◽  
Living Cells ◽  
Human Neutrophils ◽  
Protein Synthesis Inhibitor ◽  
Chemotactic Peptide ◽  
Prosthetic Group ◽  
Synthesis Inhibitor

1. A method has been developed to incorporate the apoprotein of the Ca2+-activated photoprotein obelin, and mRNA purified from the hydroid Obelia, into the cytoplasm of intact human neutrophils. This was based on internal release from pH-sensitive immunoliposomes taken up initially by phagocytosis. 2. Addition of the prosthetic group of obelin, coelenterazine, to these cells containing apo-obelin or Obelia mRNA resulted in formation of active Ca2+-activated obelin. 3. The obelin formed within the neutrophils responded to the chemotactic peptide N-formylmethionyl-leucyl-phenylalanine (1 microM) and to the membrane attack complex of complement (C5B6789n). 4. The formation of the apo-obelin from mRNA within neutrophils was inhibited by over 80% in the absence of added amino acids, and by over 90% by the protein-synthesis inhibitor puromycin (100 micrograms/ml). 5. The translation of Obelia mRNA inside cells provides a method for circumventing consumption of Ca2+-activated photoproteins during cell activation or injury, and for monitoring protein synthesis in living cells.

scholarly journals Effect of emetine, a protein synthesis inhibitor, on larval tail resorption in the ascidian Halocynthia roretzi

2006 ◽  
Vol 23 (2) ◽  
pp. 43-46
Author(s):  
Kiyotaka Matsumura ◽  
Manami Nagano ◽  
Sachiko Tsukamoto ◽  
Haruko Kato ◽  
Nobuhiro Fusetani
Keyword(s):  
Protein Synthesis ◽  
Protein Synthesis Inhibitor ◽  
Synthesis Inhibitor ◽  
Halocynthia Roretzi

scholarly journals Local protein synthesis of neuronal MT1-MMP for agrin-induced presynaptic development

Development ◽  
2021 ◽  
Vol 148 (10) ◽  
Author(s):  
Jun Yu ◽  
Marilyn Janice Oentaryo ◽  
Chi Wai Lee
Keyword(s):  
Protein Synthesis ◽  
Neuronal Development ◽  
Neuromuscular Junctions ◽  
Neuronal Cell ◽  
Time Lapse ◽  
Protein Synthesis Inhibitor ◽  
Synthesis Inhibitor ◽  
Local Protein Synthesis ◽  
Presynaptic Differentiation ◽  
Local Protein

ABSTRACT Upon the stimulation of extracellular cues, a significant number of proteins are synthesized distally along the axon. Although local protein synthesis is crucial for various stages throughout neuronal development, its involvement in presynaptic differentiation at developing neuromuscular junctions remains unknown. By using axon severing and microfluidic chamber assays, we first showed that treatment of a protein synthesis inhibitor, cycloheximide, inhibits agrin-induced presynaptic differentiation in cultured Xenopus spinal neurons. Newly synthesized proteins are prominently detected, as revealed by the staining of click-reactive cell-permeable puromycin analog O-propargyl-puromycin, at agrin bead-neurite contacts involving the mTOR/4E-BP1 pathway. Next, live-cell time-lapse imaging demonstrated the local capturing and immobilization of ribonucleoprotein granules upon agrin bead stimulation. Given that our recent study reported the roles of membrane-type 1 matrix metalloproteinase (MT1-MMP) in agrin-induced presynaptic differentiation, here we further showed that MT1-MMP mRNA is spatially enriched and locally translated at sites induced by agrin beads. Taken together, this study reveals an essential role for axonal MT1-MMP translation, on top of the well-recognized long-range transport of MT1-MMP proteins synthesized from neuronal cell bodies, in mediating agrin-induced presynaptic differentiation.

Protein synthesis requirement of the Aplysia circadian clock. Tested by active and inactive derivatives of the inhibitor anisomycin

1980 ◽  
Vol 85 (1) ◽  
pp. 33-42
Author(s):  
J. W. Jacklet
Keyword(s):  
Protein Synthesis ◽  
Circadian Clock ◽  
Compound Action Potential ◽  
Phase Shifts ◽  
Protein Synthesis Inhibitor ◽  
Synthesis Inhibitor ◽  
Inhibitor Molecule ◽  
Group 5 ◽  
Characteristic Effect ◽  
Derivatives Of

1.The circadian rhythm of compound action potential frequency recorded from the isolated eye of Aplysia in culture medium and darkness was subjected to 6 h pulse treatments with either anisomycin, a protein synthesis inhibitor, or inactive derivatives of anisomycin. 2. Anisomycin caused phase-dependent phase shifts of the rhythm as expected from previous experiments, but none of the derivative molecules caused phase shifts or perturbed the rhythm. 3. Anisomycin inhibited eye-protein synthesis by 75% at the concentrations (10(−6) M) used in the phase shifting experiments but none of the derivatives inhibited synthesis. 4. Only those molecules that actually inhibited protein synthesis caused phase shifts of the clock, although the inactive derivatives differed from anisomycin by only an acetyl group. 5. The results strengthen the conclusion that the inhibition of protein synthesis caused by anisomycin is important in perturbing the timing of the circadian clock and not some other characteristic effect of the inhibitor molecule. Together with the results from other systems, these findings imply that the daily synthesis of protein is a general requirement for circadian clocks.

A Novel Cytokine That Activates Human Neutrophils

1989 ◽  
Vol 2 (2) ◽  
pp. 63-65
Author(s):  
M. Baggiolini ◽  
A. Walz ◽  
P. Peveri ◽  
B. Dewald
Keyword(s):  
Amino Acids ◽  
Mononuclear Phagocytes ◽  
Human Neutrophils ◽  
Chemotactic Peptide ◽  
Amino Terminal ◽  
Major Form

A novel neutrophil-activating peptide was recently characterized by different laboratories and named NAF (neutrophil-activating factor) (Walz et al., 1987) MDNCF (monocyte-derived neutrophil chemotactic peptide) (Yoshimura et al., 1987) and MONAP (monocyte-derived neutrophil-activating peptide) (Gregory et al., 1988). The peptide detected in the culture fluids of stimulated human mononuclear phagocytes consists of 72 amino acids. Minor amino-terminal truncation variants are also found (Lindley et al., 1988). The major form was now obtained by recombinant methods and was shown to be biologically equivalent to purified natural NAF (Lindley et al., 1988).

scholarly journals Transforming Potential of the Herpesvirus Oncoprotein MEQ: Morphological Transformation, Serum-Independent Growth, and Inhibition of Apoptosis

1998 ◽  
Vol 72 (1) ◽  
pp. 388-395 ◽  
Author(s):  
Juinn-Lin Liu ◽  
Ying Ye ◽  
Lucy F. Lee ◽  
Hsing-Jien Kung
Keyword(s):  
Protein Synthesis ◽  
Target Genes ◽  
Rapid Development ◽  
Fibroblast Cell ◽  
Protein Synthesis Inhibitor ◽  
Morphological Transformation ◽  
Synthesis Inhibitor ◽  
Necrosis Factor Alpha ◽  
Downstream Target ◽  
T Cell Lymphomas

ABSTRACT Marek’s disease virus (MDV) induces the rapid development of overwhelming T-cell lymphomas in chickens. One of its candidate oncogenes, meq (MDV Eco Q) which encodes a bZIP protein, has been biochemically characterized as a transcription factor. Interestingly, MEQ proteins are expressed not only in the nucleoplasm but also in the coiled bodies and the nucleolus. Its novel subcellular localization suggests that MEQ may be involved in other functions beyond its transcriptional potential. In this report we show that MEQ proteins are expressed ubiquitously and abundantly in MDV tumor cell lines. Overexpression of MEQ results in transformation of a rodent fibroblast cell line, Rat-2. The criteria of transformation are based on morphological transfiguration, anchorage-independent growth, and serum-independent growth. Furthermore, MEQ is able to distend the transforming capacity of MEQ-transformed Rat-2 cells through inhibition of apoptosis. Specifically, MEQ can efficiently protect Rat-2 cells from cell death induced by multiple modes including tumor necrosis factor alpha, C2-ceramide, UV irradiation, and serum deprivation. Its antiapoptotic function requires new protein synthesis, as treatment with a protein synthesis inhibitor, cycloheximide, partially reversed MEQ’s antiapoptotic effect. Coincidentally, transcriptional induction of bcl-2 and suppression of bax are also observed in MEQ-transformed Rat-2 cells. Taken together, our results suggest that MEQ antagonizes apoptosis through regulation of its downstream target genes involved in apoptotic and/or antiapoptotic pathways.

Determinants of kinin release in isolated rat hindquarters

1998 ◽  
Vol 274 (1) ◽  
pp. R120-R125 ◽  
Author(s):  
Tohru Sakakibara ◽  
Thomas H. Hintze ◽  
Alberto Nasjletti
Keyword(s):  
Protein Synthesis ◽  
Trypsin Inhibitor ◽  
De Novo ◽  
Soybean Trypsin Inhibitor ◽  
Protein Synthesis Inhibitor ◽  
Synthesis Inhibitor ◽  
Bicarbonate Buffer ◽  
Kallikrein Inhibitor ◽  
Isolated Rat

We studied the determinants of kinin release into the venous effluent of rat hindquarters perfused with Krebs bicarbonate buffer. Kinin release in preparations perfused with control media (14.6 ± 2.5–20.7 ± 6.7 pg/15 min) was surpassed by that in preparations perfused with media containing kininase inhibitors (243 ± 53 to 276 ± 78 pg/15 min). Kinin release increased when purified kininogen (from 242 ± 43 to 3,365 ± 725 pg/15 min) or kallikrein (from 270 ± 49 to 30,649 ± 8,040 pg/15 min) was added to the perfusate. Conversely, kinin release fell when the kallikrein inhibitor aprotinin (from 272 ± 58 to 122 ± 27 pg/15 min) or soybean trypsin inhibitor (from 273 ± 52 to 195 ± 25 pg/15 min) was added. Both basal and kininogen-induced kinin release were attenuated in preparations perfused with media containing cycloheximide, a protein synthesis inhibitor, but kallikrein-induced kinin release was not. These data suggest that kinin release from perfused rat hindquarters reflects the activity of both the kinin-degrading and kinin-generating pathways and that the latter is sustained by a kallikrein manufactured de novo and by preexistent kininogen(s).

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