scholarly journals Alpha-actinin from sea urchin eggs: biochemical properties, interaction with actin, and distribution in the cell during fertilization and cleavage.

1985 ◽  
Vol 100 (2) ◽  
pp. 375-383 ◽  
Author(s):  
I Mabuchi ◽  
Y Hamaguchi ◽  
T Kobayashi ◽  
H Hosoya ◽  
S Tsukita ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Sea Urchin ◽  
Actin Filaments ◽  
Gel Filtration ◽  
Biochemical Properties ◽  
Ion Concentration ◽  
Molar Ratio ◽  
Cell Cortex ◽  
Sea Urchin Eggs ◽  
Sea Urchin Egg

A protein similar to alpha-actinin has been isolated from unfertilized sea urchin eggs. This protein co-precipitated with actin from an egg extract as actin bundles. Its apparent molecular weight was estimated to be approximately 95,000 on an SDS gel: it co-migrated with skeletal-muscle alpha-actinin. This protein also co-eluted with skeletal muscle alpha-actinin from a gel filtration column giving a Stokes radius of 7.7 nm, and its amino acid composition was very similar to that of alpha-actinins. It reacted weakly but significantly with antibodies against chicken skeletal muscle alpha-actinin. We designated this protein as sea urchin egg alpha-actinin. The appearance of sea urchin egg alpha-actinin as revealed by electron microscopy using the low-angle rotary shadowing technique was also similar to that of skeletal muscle alpha-actinin. This protein was able to cross-link actin filaments side by side to form large bundles. The action of sea urchin egg alpha-actinin on the actin filaments was studied by viscometry at a low-shear rate. It gelled the F-actin solution at a molar ratio to actin of more than 1:20, at pH 6-7.5, and at Ca ion concentration less than 1 microM. The effect was abolished by the presence of tropomyosin. Distribution of this protein in the egg during fertilization and cleavage was investigated by means of microinjection of the rhodamine-labeled protein in the living eggs. This protein showed a uniform distribution in the cytoplasm in the unfertilized eggs. Upon fertilization, however, it was concentrated in the cell cortex, including the fertilization cone. At cleavage, it seemed to be concentrated in the cleavage furrow region.

scholarly journals A 45,000-mol-wt protein-actin complex from unfertilized sea urchin egg affects assembly properties of actin.

1984 ◽  
Vol 99 (3) ◽  
pp. 994-1001 ◽  
Author(s):  
H Hosoya ◽  
I Mabuchi
Keyword(s):  
Sea Urchin ◽  
Actin Filament ◽  
Actin Polymerization ◽  
Initial Rate ◽  
Gel Filtration ◽  
Molar Ratio ◽  
Capping Protein ◽  
Sea Urchin Egg ◽  
Rate Of Polymerization ◽  
Hemicentrotus Pulcherrimus

A one-to-one complex of a 45,000-mol-wt protein and actin was purified from unfertilized eggs of the sea urchin, Hemicentrotus pulcherrimus, by means of DNase l-Sepharose affinity and gel filtration column chromatographies. Effects of the complex on the polymerization of actin were studied by viscometry, spectrophotometry, and electron microscopy. The results are summarized as follows: (a) The initial rate of actin polymerization is inhibited at a very low molar ratio of the complex to actin. (b) Acceleration of the initial rate of polymerization occurs at a relatively high, but still substoichiometric, molar ratio of the complex to actin. (c) Annealing of F-actin fragments is inhibited by the complex. (d) The complex prevents actin filaments from depolymerizing. (e) Growth of the actin filament is inhibited at the barbed end. In all cases except b, a molar ratio of less than 1:100 of the 45,000-mol-wt protein-actin complex to actin is sufficient to produce these significant effects. These results indicate that the 45,000-mol-wt protein-actin complex from the sea urchin egg regulates the assembly of actin by binding to the barbed end (preferred end or rapidly growing end) of the actin filament. The 45,000-mol-wt protein-actin complex can thus be categorized as a capping protein.

Sea urchin egg villin: identification of villin in a non-epithelial cell from an invertebrate species

1991 ◽  
Vol 100 (1) ◽  
pp. 61-71 ◽  
Author(s):  
F.S. Wang ◽  
E.M. Bonder
Keyword(s):  
Epithelial Cell ◽  
Sea Urchin ◽  
Brush Border ◽  
Actin Filaments ◽  
Binding Protein ◽  
Actin Binding ◽  
Villin Headpiece ◽  
Actin Binding Protein ◽  
Sea Urchin Eggs ◽  
Sea Urchin Egg

Fertilization of sea urchin eggs results in the rapid polymerization of actin filaments and subsequent formation of a brush border-like cortical cytoskeleton. A 110 × 10(3) Mr (110K) actin binding protein has been purified from extracts of unfertilized Strongylocentrotus purpuratus eggs. Analysis of polymerization kinetics using fluorescence and viscometry assays demonstrated that 110K accelerated the nucleation phase of actin assembly only in the presence of elevated Ca2+. The Ca(2+)-mediated effects were correlated with a decrease in sedimentable polymer and a decrease in average filament length. Addition of Ca2+ to solutions of 110K and F-actin, polymerized in the presence of EGTA, resulted in a precipitous drop in viscosity and the decreased viscosity was fully reversible upon chelation of Ca2+. The Ca2+ threshold for 110K activation was in the 10(−6) to 10(−7) M range. Nucleated assembly experiments using Limulus sperm acrosomal processes demonstrated that egg 110K capped the barbed ends of actin filaments. In the absence of Ca2+, 110K organized actin filaments into bundles at pH values less than 7.4. Anti-egg 110K antibody crossreacted with chicken intestinal epithelial cell villin and anti-porcine villin headpiece monoclonal antibody crossreacted with 110K. Further, 110K possesses an approximately 10 × 10(3) Mr terminal polypeptide segment that is immunologically related to villin headpiece. These studies demonstrate that sea urchin egg 110K is functionally, immunologically and structurally related to villin, an actin binding protein expressed in specific epithelial tissues in vertebrates. Consequently, this finding provides insight into the potential mechanisms that might determine the genesis of the cortical brush border cytoarchitecture in sea urchin eggs and further sheds light on the evolution of the villin protein family.

scholarly journals Nicotinic acid-adenine dinucleotide phosphate mobilizes Ca2+ from a thapsigargin-insensitive pool

1996 ◽  
Vol 315 (3) ◽  
pp. 721-725 ◽  
Author(s):  
Armando A. GENAZZANI ◽  
Antony GALIONE
Keyword(s):  
Nicotinic Acid ◽  
Sea Urchin ◽  
Cross Talk ◽  
Inositol Trisphosphate ◽  
Sea Urchin Eggs ◽  
Bivalent Cations ◽  
Sea Urchin Egg ◽  
Different Characteristics ◽  
Degree Of Overlap

Nicotinic acid–adenine dinucleotide phosphate (NAADP) is a novel intracellular Ca2+ releasing agent recently described in sea-urchin eggs and egg homogenates. Ca2+ release by NAADP is independent of that induced by either inositol trisphosphate (InsP3) or cyclic adenosine dinucleotide phosphate (cADPR). We now report that in sea urchin egg homogenates, NAADP releases Ca2+ from a Ca2+ pool that is distinct from those that are sensitive to InsP3 and cADPR. This organelle has distinct Ca2+ uptake characteristics: it is insensitive to thapsigargin and cyclopiazoic acid, but maintenance of the pool shows some requirement for ATP. Although the different Ca2+ pools have different characteristics, there appears to be some degree of overlap or cross-talk between the NAADP- and cADPR/InsP3-sensitive Ca2+ pools. Ca2+-induced Ca2+ release is unlikely to account for the apparent overlap between stores, since NAADP-induced Ca2+ release, in contrast with that stimulated by cADPR, is not potentiated by bivalent cations.

Ryanodine Activates Sea Urchin Eggs. (sea urchin egg/activation/ryanodine/inositol phosphate/heparin)

1992 ◽  
Vol 34 (1) ◽  
pp. 37-42 ◽  
Author(s):  
C. Sardet ◽  
I. Gillot ◽  
A. Ruscher ◽  
P. Payan ◽  
J.-P. Girard ◽  
...  
Keyword(s):  
Sea Urchin ◽  
Inositol Phosphate ◽  
Egg Activation ◽  
Sea Urchin Eggs ◽  
Sea Urchin Egg

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 Actin in triton-treated cortical preparations of unfertilized and fertilized sea urchin eggs.

1979 ◽  
Vol 82 (1) ◽  
pp. 212-226 ◽  
Author(s):  
A Spudich ◽  
J A Spudich
Keyword(s):  
Electron Microscopy ◽  
Sea Urchin ◽  
Actin Filaments ◽  
Cell Types ◽  
Optical Diffraction ◽  
Muscle Actin ◽  
Sea Urchin Eggs ◽  
Inner Surface ◽  
Low Ionic Strength ◽  
Helical Parameters

Triton-treated cortical fragments of unfertilized and fertilized sea urchin eggs prepared in the presence of greater than or equal to 5 mM EGTA contain 15-30% of the total egg actin. However, actin filaments are not readily apparent by electron microscopy on the cortical fragments of unfertilized eggs but are numerous on those of fertilized eggs. The majority of the actin associated with cortical fragments of unfertilized eggs is solubilized by dialysis against a low ionic strength buffer at pH 7.5. This soluble actin preparation (less than 50% pure actin) does not form proper filaments in 0.1 M KCl and 3 mM MgCl2, whereas actin purified from this preparation does, as judged by electron microscopy. Optical diffraction analysis reveals that these purified actin filaments have helical parameters very similar to those of muscle actin. Furthermore, the properties of the purified actin with regard to activation of myosin ATPase are similar to those of actin from other cell types. The possibility that actin is maintained in a nonfilamentous form on the inner surface of the unfertilized egg plasma membrane and is induced to assemble upon fertilization is discussed.

Production of Novel Exopolysaccharide with Emulsifying Ability from Marine Microorganism, Alteromonas sp. Strain 00SS11568

2005 ◽  
Vol 277-279 ◽  
pp. 155-161 ◽  
Author(s):  
Joung Han Yim ◽  
Se Hun Ahn ◽  
Sung Jin Kim ◽  
Yoo Kyung Lee ◽  
Kyu Jin Park ◽  
...  
Keyword(s):  
Inorganic Compounds ◽  
Gel Filtration ◽  
Optimal Growth ◽  
Growth Conditions ◽  
Biochemical Properties ◽  
Medium Composition ◽  
Molar Ratio ◽  
Bacterial Strains ◽  
Average Molecular Mass ◽  
Emulsifying Ability

To find a novel exopolysaccharide, marine bacterial strains were isolated from coastal regions of Korea. Strain 00SS11568 was then selected as it produced a mucous exopolysaccharide during the stationary phase in a batch culture. The isolate was identified as Alteromonas sp. based on its 16S rDNA sequence, morphological, and biochemical properties. The exopolysaccharide, designated as p-11568, exhibited an emulsifying ability. The Emulsification Index (E24) of 0.1% p- 11568 was 77.4% with an emulsified kerosene content, and was higher than those of commercial polysaccharides, such as xanthan gum (26.1%), gellan gum (1.3%), and sodium alginate (2.0%). p- 11568 was found to be composed of glucose and galactose as the main natural sugars in a molar ratio of 1.3:1, along with uronic acid (18.9%, w/w) and sulfate groups (1.2% w/w). The average molecular mass was 4.4 x 105 daltons by gel filtration chromatography. The effects of pH, temperature, inorganic compounds, and C and N sources were tested to obtain the optimal medium composition for the production of p-11568. Under optimal growth conditions with the M-11568 medium, 14.9 g of crude p-11568 per liter was obtained.

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