scholarly journals Sea urchin tube feet: unique structures that allow a cytological and molecular approach to the study of actin and its gene expression.

1981 ◽  
Vol 89 (1) ◽  
pp. 109-114 ◽  
Author(s):  
J Kabat-Zinn ◽  
R H Singer
Keyword(s):  
Sea Urchin ◽  
Gel Electrophoresis ◽  
Messenger Rna ◽  
Strongylocentrotus Purpuratus ◽  
Strongylocentrotus Droebachiensis ◽  
Blue Staining ◽  
Phage Library ◽  
Two Dimensional ◽  
Arbacia Punctulata ◽  
Tube Feet

Actin is the major extractable protein component from the tube feet of four different species of sea urchin: Arbacia punctulata, Strongylocentrotus purpuratus, Strongylocentrotus droebachiensis, and Diadema setosum. Actin made up as much as 60% of the total Coomassie Blue-staining material after SDS polyacrylamide gel electrophoresis and densitometer analysis. Two-dimensional gel electrophoresis resolved two, and possible three, species of actin for each sea urchin of which the dominant component was analogous to the beta form in vertebrates. In a cell-free system from rabbit reticulocytes, total RNA from tube feet stimulated the synthesis of one protein that represented 80% of the total methionine incorporation, migrated with the properties characteristic of actin in a two-dimensional gel system, and on proteolysis yielded fragments identical to purified rabbit actin. The mRNAs from the tube feet of two divergent species of sea urchin, Arbacia punctulata and Strongylocentrotus purpuratus, synthesized actins differing by less than 0.02 pH unit for each isospecies 90% of the DNA copied from tube foot RNA by reverse transcriptase represented a highly abundant sequence class judged by copy DNA(cDNA)-RNA excess hybridization. At least two-thirds of this class represented a low-complexity component, with a Rot1/2 about three times that expected for actin messenger RNA. The remarkable degree of conservation of the actin protein is reflected in concomitant conservation of the protein-coding nucleotide sequences of the messenger RNA, which has allowed the use of a cDNA probe to isolate actin sequences from a human phage library.

scholarly journals Striated flagellar roots: isolation and partial characterization of a calcium-modulated contractile organelle.

1984 ◽  
Vol 99 (3) ◽  
pp. 962-970 ◽  
Author(s):  
J L Salisbury ◽  
A Baron ◽  
B Surek ◽  
M Melkonian
Keyword(s):  
Gel Electrophoresis ◽  
Calcium Binding ◽  
Protein Band ◽  
Blue Staining ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Flagellar Roots ◽  
Beta Form ◽  
Cell Extracts ◽  
Tetraselmis Striata

We report the isolation of striated flagellar roots from the Prasinophycean green alga Tetraselmis striata using sedimentation in gradients of sucrose and flotation on gradients of colloidal silica. PAGE in the presence of 0.1% SDS demonstrates that striated flagellar roots are composed of a number of polypeptides, the most predominant one being a protein of 20,000 Mr. The 20,000 Mr protein band represents approximately 63% of the Coomassie Brilliant Blue staining of gels of isolated flagellar roots. Two-dimensional gel electrophoresis (isoelectric focusing and SDS PAGE) resolves the major 20,000 Mr flagellar root protein into two components of nearly identical Mr, but of differing isoelectric points (i.e., pl's of 4.9 and 4.8), which we have designated 20,000-Mr-alpha and 20,000-Mr-beta, respectively. Densitometric scans of two-dimensional gels of cell extracts indicate that the 20,000-Mr-alpha and -beta polypeptides vary, in their stoichiometry, between 2:1 and 1:1. This variability appears to be related to the state of contraction or extension of the striated flagellar roots at the time of cell lysis. Incubation of cells with 32PO4 followed by analysis of cell extracts by two-dimensional gel electrophoresis and autoradiography reveals that the more acidic 20,000-Mr-beta component is phosphorylated and the 20,000-Mr-alpha component contains no detectable label. These results suggest that the 20,000-Mr-alpha component is converted to the more acidic 20,000-Mr-beta form by phosphorylation. Both the 20,000-Mr-alpha and -beta flagellar root components exhibit a calcium-induced reduction in relative electrophoretic mobilities in two-dimensional alkaline urea gels. Antiserum raised in rabbits against the 20,000-Mr protein binds to both the 20,000-Mr-alpha and 20,000-Mr-beta forms of the flagellar root protein when analyzed by electrophoretic immunoblot techniques. Indirect immunofluorescence on vegetative or interphase cells demonstrate that the antibodies bind to two cyclindrical organelles located in the anterior region of the cell. Immunocytochemical investigations at ultrastructural resolution using this antiserum and a colloidal gold-conjugated antirabbit-IgG reveals immunospecific labeling of striated flagellar roots and their extensions. We conclude that striated flagellar roots are simple ion-sensitive contractile organelles composed predominantly of a 20,000 Mr calcium-binding phosphoprotein, and that this protein is largely responsible for the motile behavior of these organelles.

scholarly journals Identification and characterization of glycoproteins after extraction of bovine chromaffin-granule membranes with lithium di-iodosalicylate. Purification of glycoprotein II from the soluble fraction

1990 ◽  
Vol 270 (1) ◽  
pp. 57-61 ◽  
Author(s):  
D L Christie ◽  
D J Palmer
Keyword(s):  
Gel Electrophoresis ◽  
Soluble Fraction ◽  
Secretory Granules ◽  
Insoluble Fraction ◽  
Blue Staining ◽  
Two Dimensional ◽  
Membrane Glycoproteins ◽  
Chromaffin Granule ◽  
Two Dimensional Gel Electrophoresis ◽  
Coomassie Blue Staining

Chromaffin-granule membranes were separated into insoluble and soluble fractions after extraction with lithium di-iodosalicylate (LDIS). These fractions were characterized by one- and two-dimensional gel electrophoresis, and glycoproteins were detected after electroblotting with peroxidase-labelled concanavalin A and wheat-germ agglutinin (WGA). The LDIS-insoluble fraction contained components identified as glycoproteins III, H, J and K (carboxypeptidase H). Microsequence analysis indicated that component J is an N-terminally extended form of glycoprotein K. A major glycoprotein, GpII (Mr 80,000-100,000), present in the LDIS-soluble fraction was purified by affinity chromatography on WGA-Sepharose. This was characterized by one- and two-dimensional gel electrophoresis with Coomassie Blue staining, by amino acid analysis and automated N-terminal sequence analysis. Extraction of chromaffin-granule membranes with LDIS is a simple and rapid procedure that facilitates studies concerned with the structure and function of membrane glycoproteins from these and other secretory granules.

Experimental determinations of tubulin in the in vivo mitotic apparatus of sea urchin zygotes

1980 ◽  
Vol 58 (11) ◽  
pp. 1277-1285 ◽  
Author(s):  
Arthur Forer ◽  
D. E. Larson ◽  
A. M. Zimmerman
Keyword(s):  
Sea Urchin ◽  
Gel Electrophoresis ◽  
Dry Matter ◽  
Strongylocentrotus Purpuratus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Mitotic Apparatus ◽  
Cell Lysate ◽  
Hexylene Glycol

Mitotic apparatus (MA) were isolated from zygotes of a sea urchin (Strongylocentrotus purpuratus), using hexylene glycol (pH 6.4) as lysing–stabilizing agent. Protein was measured in the MA pellet and in the remainder of the cell lysate (using the Lowry procedure). Tubulin was measured in the MA pellet and in the remainder of the cell lysate (using microdensitometry of stained gels after sodium dodecyl sulphate – polyacrylamide gel electrophoresis). From these data we calculated the maximum possible amounts of tubulin in the isolated MA and in the MA in vivo; in these calculations we assumed that all the tubulin in the cell is associated with the MA, and we assumed that, as reported in the literature, the MA lose 90% of their dry matter during the isolation. We conclude that tubulin probably comprises less than 7% of the protein in the in vivo MA, and, even if there are very large errors, tubulin is considerably less than haf the protein in the MA.

Neurosensory and neuromuscular organization in tube feet of the sea urchin Strongylocentrotus purpuratus

2011 ◽  
Vol 519 (17) ◽  
pp. 3566-3579 ◽  
Author(s):  
Cavit Agca ◽  
Milad C. Elhajj ◽  
William H. Klein ◽  
Judith M. Venuti
Keyword(s):  
Sea Urchin ◽  
Strongylocentrotus Purpuratus ◽  
Tube Feet

scholarly journals Identification of a sperm receptor on the surface of the eggs of the sea urchin Arbacia punctulata.

1977 ◽  
Vol 72 (1) ◽  
pp. 35-46 ◽  
Author(s):  
E Schmell ◽  
B J Earles ◽  
C Breaux ◽  
W J Lennarz
Keyword(s):  
Sea Urchin ◽  
Concanavalin A ◽  
Strongylocentrotus Purpuratus ◽  
Soluble Form ◽  
Ionophore A23187 ◽  
Soluble Receptor ◽  
Specific Receptor ◽  
Marked Contrast ◽  
Arbacia Punctulata ◽  
Species Specific

The possibility that the surface of the egg of the sea urchin Arbacia punctulata contains a species-specific receptor for sperm has been investigated. The extent of fertilization of eggs of A. punctulata, which is proportional to the number of sperm, is unaffected by the presence of either eggs or membranes prepared from eggs of Strongylocentrotus purpuratus. In marked contrast, membranes prepared from eggs of A. punctulata quantitatively inhibit fertilization of A. punctulata eggs by A. punctulata sperm. Several lines of evidence indicate that this inhibition is due to the presence of a membrane-associated glycoprotein that binds to the sperm, thus preventing them from interacting with receptor on the surface of the eggs. First, eggs treated with trypsin are incapable of being fertilized, although they can be activated with the Ca2+ ionophore A23187. Moreover, membranes prepared from eggs pretreated with trypsin do not inhibit fertilization of eggs. Second, receptor isolated in soluble form from surface membranes binds to sperm and thus prevents them from fertilizing eggs; the inhibition by soluble receptor is species-specific. Third, the soluble receptor binds to concanavalin A-Sepharose. Fourth, eggs are incapable of being fertilized if they are pretreated with concanavalin A. The specificity of inhibition, and the affect of trypsin and concanavalin A on intact eggs, suggest that the receptor is a species-specific macromolecule located on the surface of the eggs. The sensitivity of the receptor to trypsin, and its ability to bind to concanavalin A, indicate that it is a glycoprotein.

scholarly journals Adhesive plasticity among populations of purple sea urchin (Strongylocentrotus purpuratus)

2020 ◽  
Vol 223 (15) ◽  
pp. jeb228544
Author(s):  
Alyssa Y. Stark ◽  
Carla A. Narvaez ◽  
Michael P. Russell
Keyword(s):  
Sea Urchin ◽  
Sea Urchins ◽  
Strongylocentrotus Purpuratus ◽  
Population Level ◽  
Adhesive System ◽  
Adhesive Force ◽  
Artificial Substrates ◽  
Wave Forces ◽  
Skeletal Morphology ◽  
Tube Feet

ABSTRACTSea urchins native to the nearshore open coast experience periods of high, repeated wave forces that can result in dislodgement. To remain attached while clinging and locomoting across rocky substrates, sea urchins use adhesive tube feet. Purple sea urchins (Strongylocentrotus purpuratus) adhere to a variety of rock substrates (e.g. sandstone, mudstone, granite), and display morphological plasticity (skeletal morphology) to native substrate. We tested the hypothesis that their adhesive system is also plastic and varies as a function of native population and substrate. The results of our study support our hypothesis. Sea urchins from sandstone adhere less strongly to most substrates than those native to mudstone and granite rock. Sandstone produced the lowest whole animal adhesive force values across all populations, suggesting that this rock type is particularly challenging for sea urchins to adhere to. The number of adhesive tube feet that failed during experimental trials and the area used by sea urchins to attach, matches closely with whole animal adhesive force values: higher forces resulted in more tube foot failure and larger attachment area. On artificial substrates (glass and Plexiglass), differences in adhesion among populations was consistent with differences in adhesion on rock substrates except on glass, where sea urchins native to sandstone adhered more strongly to glass than any other substrate tested. To our knowledge, this study is the first to describe population-level plasticity in a biological adhesive system related to native substrate, and has significant implications for sea urchin ecology, behavior and functional morphology.

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