scholarly journals THE EFFECTS OF CARBON MONOXIDE INHIBITION ON ATP LEVEL AND THE RATE OF MITOSIS IN THE SEA URCHIN EGG

1963 ◽  
Vol 17 (2) ◽  
pp. 315-319 ◽  
Author(s):  
David Epel
Keyword(s):  
Carbon Monoxide ◽  
Sea Urchin ◽  
Strongylocentrotus Purpuratus ◽  
Specific Inhibitor ◽  
Atp Synthesis ◽  
Luciferase Assay ◽  
Basic Premise ◽  
Atp Level ◽  
Sea Urchin Egg ◽  
Kinetics Of

The requirements for ATP synthesis during the various phases of mitosis were investigated in the oxygen-requiring eggs of the sea urchin, Strongylocentrotus purpuratus. CO in the dark, a specific inhibitor of respiration, was used to inhibit ATP synthesis. The kinetics of respiratory inhibition were determined by analyzing ATP levels with the luciferin-luciferase assay. The kinetics of mitotic inhibition were determined by analysis of the rate of mitosis. It was found that CO inhibition resulted in a decrease in the normal ATP level. Coincident with this decrease was a decrease in the rate of mitosis which stops completely when the ATP drops below 50 per cent of the normal level. With the use of various degrees of CO inhibition, the rate of mitosis is shown to be related to the resultant ATP level. These results contradict the basic premise of the energy reservoir hypothesis, and also disagree with other reports that cells in mitosis are insensitive to inhibitors of energy metabolism. Data are presented which demonstrate that these conflicting reports result from insufficient inhibition of ATP synthesis. The above findings all indicate that mitosis depends on the continuous synthesis and utilization of ATP.

The Respiratory and Glycolytic Enzymes of Sea-Urchin Eggs

1954 ◽  
Vol 31 (2) ◽  
pp. 208-217
Author(s):  
MARTYNAS YČAS
Keyword(s):  
Cytochrome C ◽  
Sea Urchin ◽  
Strongylocentrotus Purpuratus ◽  
Lactic Dehydrogenase ◽  
Glycolytic Pathway ◽  
Endogenous Respiration ◽  
Centrifugal Field ◽  
Lytechinus Pictus ◽  
Sea Urchin Eggs ◽  
Sea Urchin Egg

1. Activity corresponding to phosphoglucomutase, phosphohexoisomerase, aldolase, triosephosphate dehydrogenase, enolase and lactic dehydrogenase has been demonstrated in homogenates prepared from unfertilized sea-urchin eggs (Strongylocentrotus purpuratus and Lytechinus pictus). 2. The presence of cytochromes a and b1 has been confirmed. These cytochromes sediment in a relatively low centrifugal field. 3. No cytochrome c could be demonstrated, although cytochrome c is both reduced and oxidized by homogenates, and addition of cytochrome c increases the endogenous respiration and oxidation of succinate. 4. These results support the view that the usual glycolytic pathway operates in the sea-urchin egg and is the principal route of oxidation of carbohydrate.

scholarly journals Hierarchies of protein cross-linking in the extracellular matrix: involvement of an egg surface transglutaminase in early stages of fertilization envelope assembly.

1988 ◽  
Vol 107 (6) ◽  
pp. 2447-2454 ◽  
Author(s):  
D E Battaglia ◽  
B M Shapiro
Keyword(s):  
Cell Surface ◽  
Sea Urchin ◽  
Physiological Function ◽  
Strongylocentrotus Purpuratus ◽  
Cross Linking ◽  
Early Step ◽  
Transglutaminase Activity ◽  
Sea Urchin Egg ◽  
A Cell ◽  
Secretory Products

The involvement of transglutaminase activity in fertilization envelope (FE) formation was investigated using eggs from the sea urchin, Strongylocentrotus purpuratus. Eggs fertilized in the presence of the transglutaminase inhibitors, putrescine and cadaverine, had disorganized and expanded FEs with inhibition of the characteristic I-T transition. The permeability of the FE was increased by these agents, as revealed by the loss of proteins from the perivitelline space and the appearance of ovoperoxidase activity in supernates from putrescine-treated eggs. [3H]putrescine was incorporated into the FE during fertilization in a reaction catalyzed by an egg surface transglutaminase that could also use dimethylcasein as a substrate in vitelline layer-denuded eggs. Egg secretory products alone had no transglutaminase activity. The cell surface transglutaminase activity was transient and maximal within 4 min of activation. The enzyme was Ca2+ dependent and was inhibited by Zn2+. We conclude that sea urchin egg surface transglutaminase catalyzes an early step in a hierarchy of cross-linking events during FE assembly, one that occurs before ovoperoxidase-mediated dityrosine formation (Foerder, C. A., and B. M. Shapiro. 1977. Proc. Natl. Acad. Sci. USA. 74:4214-4218). Thus it provides a graphic example of the physiological function of a cell surface transglutaminase.

scholarly journals Unique kinetics of nicotinic acid–adenine dinucleotide phosphate (NAADP) binding enhance the sensitivity of NAADP receptors for their ligand

2000 ◽  
Vol 352 (3) ◽  
pp. 725-729 ◽  
Author(s):  
Sandip PATEL ◽  
Grant C. CHURCHILL ◽  
Antony GALIONE
Keyword(s):  
Nicotinic Acid ◽  
Sea Urchin ◽  
Binding Sites ◽  
Cell Types ◽  
Unique Property ◽  
Single Class ◽  
High Affinity ◽  
Sea Urchin Egg ◽  
Low Concentrations ◽  
Kinetics Of

Nicotinic acidŐadenine dinucleotide phosphate (NAADP) is a novel and potent Ca2+-mobilizing agent in sea urchin eggs and other cell types. Little is known, however, concerning the properties of the putative intracellular NAADP receptor. In the present study we have characterized NAADP binding sites in sea urchin egg homogenates. [32P]NAADP bound to a single class of high-affinity sites that were reversibly inhibited by NaCl but insensitive to pH and Ca2+. Binding of [32P]NAADP was lost in preparations that did not mobilize Ca2+ in response to NAADP, indicating that [32P]NAADP probably binds to a receptor mediating Ca2+ mobilization. Addition of excess unlabelled NAADP, at various times after initiation of [32P]NAADP binding, did not result in displacement of bound [32P]NAADP. These data show that NAADP becomes irreversibly bound to its receptor immediately upon association. Accordingly, incubation of homogenates with low concentrations of NAADP resulted in maximal labelling of NAADP binding sites. This unique property renders NAADP receptors exquisitely sensitive to their ligand, thereby allowing detection of minute changes in NAADP levels.

scholarly journals Colchicine-binding activity distinguishes sea urchin egg and outer doublet tubulins.

1984 ◽  
Vol 99 (1) ◽  
pp. 37-41 ◽  
Author(s):  
L Wilson ◽  
H P Miller ◽  
T A Pfeffer ◽  
K F Sullivan ◽  
H W Detrich
Keyword(s):  
Sea Urchin ◽  
Strongylocentrotus Purpuratus ◽  
Binding Activity ◽  
Binding Affinities ◽  
Regulatory Site ◽  
Colchicine Binding Site ◽  
Sea Urchin Egg ◽  
Enthalpy And Entropy Changes ◽  
Enthalpy And Entropy ◽  
Significant Chemical

The colchicine-binding activity of tubulin has been utilized to distinguish the tubulins from two distinct microtubule systems of the same species, the sea urchin Strongylocentrotus purpuratus. We have analyzed the colchicine-binding affinities of highly purified tubulins from the unfertilized eggs and from the flagellar outer doublet microtubules by van't Hoff analysis, and have found significant differences in the free energy, enthalpy, and entropy changes characterizing the binding of colchicine to the two tubulins. The data indicate that significant chemical differences in the tubulins from the two functionally distinct microtubule systems exist, and that the differences are expressed in the native forms of the tubulins. Our findings are discussed in terms of the possibility that the colchicine-binding site may be an important regulatory site on the tubulin molecule.

scholarly journals Ca2+ release triggered by nicotinate adenine dinucleotide phosphate in intact sea urchin eggs

1995 ◽  
Vol 312 (3) ◽  
pp. 955-959 ◽  
Author(s):  
C M Perez-Terzic ◽  
E N Chini ◽  
S S Shen ◽  
T P Dousa ◽  
D E Clapham
Keyword(s):  
Sea Urchin ◽  
Specific Inhibitor ◽  
Intact Cells ◽  
Sea Urchin Eggs ◽  
Sea Urchin Egg ◽  
Cyclic Adp Ribose ◽  
Inositol 1,4,5 Trisphosphate ◽  
Dose Dependent Increase ◽  
Dose Dependent

Nicotinate adenine dinucleotide phosphate (NAADP) was recently identified [Lee and Aarhus (1995) J. Biol. Chem. 270, 2152-2157; Chini, Beers and Dousa (1995) J. Biol. Chem. 270, 3116-3223] as a potent Ca(2+)-releasing agent in sea urchin egg homogenates. NAADP triggered Ca2+ release by a mechanism that was distinct from inositol 1,4,5-trisphosphate (InsP3)- and cyclic ADP-ribose (cADPR)-induced Ca2+ release. When NAADP was microinjected into intact sea urchin eggs it induced a dose-dependent increase in cytoplasmic free Ca2+ which was independent of the extracellular [Ca2+]. The Ca2+ waves elicited by microinjections of NAADP originated at the site of injection and swept across the cytosol. As previously found in sea urchin egg homogenates, NAADP-induced Ca2+ release in intact eggs was not blocked by heparin or by prior desensitization to InsP3 or cADPR. Thio-NADP, a specific inhibitor of the NAADP-induced Ca2+ release in sea urchin homogenates [Chini, Beers and Dousa (1995) J. Biol. Chem. 270, 3116-3223] blocked NAADP (but not InsP3 or cADPR) injection-induced Ca2+ release in intact sea urchin eggs. Finally, fertilization of sea urchin eggs abrogated subsequent NAADP-induced Ca2+ release, suggesting that the NAADP-sensitive Ca2+ pool may participate in the fertilization response. This study demonstrates that NAADP acts as a selective Ca(2+)-releasing agonist in intact cells.

Phosphate-Transport Appearance in the Sea-Urchin Egg. II. Analysis of the Genome Function with Centrifuged Eggs and Ethidium Bromide

1974 ◽  
Vol 52 (6) ◽  
pp. 1186-1191
Author(s):  
Joël de la Noüe
Keyword(s):  
Ethidium Bromide ◽  
Sea Urchin ◽  
Phosphate Uptake ◽  
Strongylocentrotus Purpuratus ◽  
Nuclear Genome ◽  
Phosphate Transport ◽  
Carrier System ◽  
Purple Sea Urchin ◽  
Sea Urchin Egg ◽  
Phosphate Carrier

Ethidium bromide, an inhibitor of mitochondrial transcription, inhibits phosphate uptake, valine incorporation, and uridine incorporation in fertilized eggs of Strongylocentrotus purpuratus, the purple sea urchin. Phosphate uptake, valine incorporation, and valine influx are also inhibited by this drug in artificially activated nucleate and anucleate egg fragments. Ethidium bromide does not affect the egg respiration and it has no measurable effect on ATP level and labeling. It is concluded that the post-fertilization appearance of the phosphate-carrier system does not require the participation of the nuclear genome but that of the mitochondrial one. It is likely that the same proposal holds for the L-valine-transport system. Some effects of ethidium bromide on development are exposed.

The GTP-binding protein RhoA localizes to the cortical granules of Strongylocentrotus purpuratus sea urchin egg and is secreted during fertilization

2000 ◽  
Vol 79 (2) ◽  
pp. 81-91 ◽  
Author(s):  
Patricia Cuéllar-Mata ◽  
Guadalupe Martínez-Cadena ◽  
Juana López-Godínez ◽  
Armando Obregón ◽  
Jesús García-Soto
Keyword(s):  
Sea Urchin ◽  
Binding Protein ◽  
Strongylocentrotus Purpuratus ◽  
Cortical Granules ◽  
Gtp Binding Protein ◽  
Gtp Binding ◽  
Sea Urchin Egg

scholarly journals Metabolic similarities between fertilization and phagocytosis. Conservation of a peroxidatic mechanism.

1979 ◽  
Vol 149 (4) ◽  
pp. 938-953 ◽  
Author(s):  
S J Klebanoff ◽  
C A Foerder ◽  
E M Eddy ◽  
B M Shapiro
Keyword(s):  
Sea Urchin ◽  
Polymorphonuclear Leukocytes ◽  
Cortical Granules ◽  
Sea Urchin Eggs ◽  
Sea Urchin Egg ◽  
Hatching Process ◽  
Fertilization Membrane ◽  
Catalyzed Reactions ◽  
Kinetics Of ◽  
Estrogen Binding

At the time of fertilization, sea urchin eggs release a peroxidase which, together with H2O2 generated by a respiratory burst, is responsible for hardening of the fertilization membrane. We demonstrate here that the ovoperoxidase of unfertilized eggs is located in cortical granules and, after fertilization, is concentrated in the fertilization membrane. Fertilization of sea urchin eggs or their parthenogenetic activation with the ionophor A23187 also results in (a) the conversion of iodide to a trichloroacetic acid-precipitable form (iodination), (b) the deiodination of eggs exogenously labeled with myeloperoxidase and H2O2, (c) the degradation of thyroxine as measured by the recovery of the released radioiodine at the origin and in the inorganic iodide spot on paper chromatography, and (d) the conversion of estradiol to an alcohol-precipitable form (estrogen binding). The iodination reaction and the binding of estradio occurs predominantly in the fertilization membrane where the ovoperoxidase is concentrated. From the estimation of the kinetics of incorporation of iodine, we determine that the peroxidative system is active for 30 min after fertilization, long after hardening of the fertilization membrane is complete. Most of the bound iodine is lost during the hatching process. Iodination of albumin is catalyzed by the material released from the egg during fertilization, when combined with H2O2 and iodide. Iodination, thyroxine degradation, and estradiol binding are inhibited by azide, cyanide, aminotriazole, methimazole, ascorbic acid and ergothioneine, all of which can inhibit peroxidase-catalyzed reactions. These responses of the sea urchin egg to fertilization are strikingly similar to the changes induced in polymorphonuclear leukocytes by phagocytosis and, in both instances, a peroxidative mechanism may be involved.

scholarly journals Monoclonal antibodies to dynein subunits reveal the existence of cytoplasmic antigens in sea urchin egg.

1984 ◽  
Vol 98 (5) ◽  
pp. 1842-1850 ◽  
Author(s):  
G Piperno
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Sea Urchin ◽  
Strongylocentrotus Purpuratus ◽  
Apparent Molecular Weight ◽  
Mitotic Apparatus ◽  
Cellular Fraction ◽  
Sea Urchin Egg ◽  
Monospecific Antibodies

Monoclonal antibodies directed against subunits of a sea urchin flagellar dynein were used to test for the presence of cytoplasmic antigens in preparations of fertilized eggs and mitotic apparati . A 9-10 S complex composed of 330,000-, 134,000-, and 126,000-mol-wt subunits was isolated from outer arms of Strongylocentrotus purpuratus sperm flagella and used to characterize the antibodies. Seven monospecific antibodies to the 330,000 subunit and two against the 134,000 subunit of the 9-10 S complex were identified by binding to nitrocellulose blots of electrophoretograms resolving polypeptides from different dynein preparations. The antibodies were applied also to blots of polypeptides from fertilized sea urchin egg at the first metaphase and a cellular fraction of mitotic apparati . Three of the antibodies to the 330,000 subunit bound to a cytoplasmic polypeptide of approximately the same molecular weight and the two antibodies to the smaller subunits recognized a polypeptide of 124,000 apparent molecular weight. Both antigens appeared to be enriched in the fraction containing mitotic apparati . These results indicate that polypeptides similar to two subunits of the 9-10 S complex are present in eggs at metaphase, and they are apparently associated with the mitotic apparatus.

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