scholarly journals Biochemical and cytochemical evidence for ATPase activity in basal bodies isolated from oviduct.

1977 ◽  
Vol 74 (2) ◽  
pp. 547-560 ◽  
Author(s):  
R G Anderson
Keyword(s):  
Enzyme Activity ◽  
Atpase Activity ◽  
Basal Body ◽  
Divalent Cation ◽  
Specific Activity ◽  
Basal Bodies ◽  
Ph Optimum ◽  
Outer Portion ◽  
Chicken Oviduct ◽  
Basal Foot

Biochemical and cytochemical techniques were used to determine whether oviduct basal bodies have ATPase activity. All studies were carried out on basal bodies isolated and purified from the chicken oviduct. These preparations contained structurally intact basal bodies with basal feet, rootlet, and alar sheet accessory structures. Whereas the specific activity of the basal body ATPase in 2 mM Ca++ or 2 mM Mg++, 1 mM ATP, pH 8.0, averaged 0.04 mumol Pi/min per mg protein, higher concentrations of either cation inhibited the enzyme activity. Furthermore, the pH optimum for this reaction was pH 8.5. In comparison, the ATPase activity in cilia purified and measured under conditions identical to those for determining the basal body ATPase activity averaged 0.07 mumol Pi/min per mg protein. However, the activity increased at higher concentrations of divalent cation, and the pH optimum was pH 10.0. By cytochemical procedures for localizing ATPase activity, ATP-dependent reaction product in isolated basal bodies was found to be confined to: (a) the cross-striations of the rootlet; (b) the outer portion of the basal foot; (c) the alar sheets; and (d) the triplet microtubules. It is concluded that basal bodiesve an intrinsic ATPase activity that, by a variety of criteria, can be distinguished from the ATPase activity found in cilia.

Phosphoglycollate Phosphatase in Marine Algae: Isolation and Characterization from Halimeda cylindracea

1976 ◽  
Vol 3 (1) ◽  
pp. 105 ◽  
Author(s):  
DD Randall
Keyword(s):  
Divalent Cation ◽  
Marine Algae ◽  
Marine Alga ◽  
Specific Activity ◽  
Ph Optimum ◽  
Isolation And Characterization

Phosphoglycollate phosphatase was partially purified (107-fold) from the marine alga H. cylindracea. The partially purified enzyme was almost completely specific for phosphoglycollate as substrate, and had the following properties: a specific activity of 2.73, an apparent Km for phosphoglycollate of 8 x 10-4M, a pH optimum of 7.5 - 8 and a divalent cation requirement. The presence of the enzyme is discussed in relation to photorespiration in this alga.

UDP-Glucose: Flavonol 7-O-glucosyltransferase Activity in Flower Extracts of Chrysanthemum segetum

1997 ◽  
Vol 52 (3-4) ◽  
pp. 153-158 ◽  
Author(s):  
K. Stich ◽  
H. Halbwirth ◽  
F. Wurst ◽  
G. Forkmann
Keyword(s):  
Enzyme Activity ◽  
Specific Activity ◽  
Temperature Stability ◽  
Inhibitory Effect ◽  
Hydroxyl Group ◽  
Ph Optimum ◽  
Yellow Colour ◽  
Commercial Strain ◽  
Strong Inhibitory Effect ◽  
Chrysanthemum Segetum

Abstract The yellow colour of Chrysanthemum segetum petals is due to the presence of the 7-O-glucosides of quercetin and particularly gossypetin (8-hydroxyquercetin). In petal extracts of C. segetum an enzyme was demonstrated which catalyzes the transfer of the glucosyl moiety of uridine 5'-diphosphoglucose (UDPG) to the 7-hydroxyl group of flavonols with gossypetin and quercetin as the best substrates. Besides flavonols flavanones and flavones were found to be glucosylated in the 7-position. The pH-optimum of the reaction highly depended on the substrate used. With quercetin as substrate, maximal enzyme activity occurred at a pH of 8.25 and a temperature of 25 °C, but 7-O-glucosylation also proceeded at low temperatures. Studies on temperature stability revealed, that there was no influence on the glucosylation reaction up to 40 °C. Higher temperatures led to a loss of enzyme activity. Using gossypetin as a substrate a similar course of temperature stability was observed. Addition of Mg2+, Ca2+ and KCN slightly stimulated 7-O-glucosylation, whereas Co2+, Cu2+, Fe2+, Hg2+, p-hydroxymercuribenzoate and N-ethylmaleimide showed a strong inhibitory effect. Additional enzymatic studies were performed with the commercial strain " Stern des Orients" where gossypetin 7-O-glucoside is restricted to the inner parts of the petals. For enzyme extracts from both parts of the petals gossypetin was found to be the most attractive substrate. In comparison to quercetin (133.4 μkat / kg protein) an about three times higher specific activity of the 7-O-glucosyltransferase(s) was determined with gossypetin (382.1 μkat/ kg protein) as substrate, indicating that hydroxylation of quercetin in 8-position to gossypetin precedes 7-O-glucosylation.

The soluble adenosine triphosphatase of Thiobacillus ferrooxidans

1975 ◽  
Vol 21 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Catherine Adapoe ◽  
Marvin Silver
Keyword(s):  
Enzyme Activity ◽  
Atpase Activity ◽  
Gel Electrophoresis ◽  
Inorganic Phosphate ◽  
Adenosine Triphosphatase ◽  
Thiobacillus Ferrooxidans ◽  
Polyacrylamide Gel Electrophoresis ◽  
Histochemical Analysis ◽  
Ph Optimum ◽  
Major Band

Adenosine triphosphatase (ATPase) from Thiobacilhis ferrooxidans was purified 55-fold. Polyacrylamide gel electrophoresis of the most purified fraction showed only one major band; histochemical analysis showed that the ATPase activity was associated with this band. The pH optimum is 9–10. The enzyme hydrolyzed ATP stoichiometrically to ADP and inorganic phosphate, the Km for this substrate being 7.75 × 10−3 M. GTP and ITP are alternate substrates, the Km values for these being 6.71 × 10−3 M and 3.12 × 10−3 M, respectively. ADP is slightly hydrolyzed. Magnesium, manganese, and calcium can serve as cofactors; Km values for these are 2.0 × 10−3 M, 9.4 × 10−4 M, and 8.0 × 10−4 M, respectively. The enzyme activity was not activated by either sodium or potassium, but a combination of the two ions were inhibitory. Azide and p-hydroxymercuribenzoate strongly inhibited the enzyme activity, whereas cyanide, dinitrophenol, and N, N′-dicyclohexylcarbodiimide (DCCD) were without effect. The enzyme was cold labile at 0 °C, but was more stable at 18–24 °C.

scholarly journals THE FORMATION OF BASAL BODIES (CENTRIOLES) IN THE RHESUS MONKEY OVIDUCT

1971 ◽  
Vol 50 (1) ◽  
pp. 10-34 ◽  
Author(s):  
Richard G. W. Anderson ◽  
Robert M. Brenner
Keyword(s):  
Rhesus Monkey ◽  
Basal Body ◽  
Wall Material ◽  
Basal Bodies ◽  
Major Pathway ◽  
Tubule Formation ◽  
Body Formation ◽  
Body Production ◽  
Basal Foot ◽  
Section Analysis

Basal body replication during estrogen-driven ciliogenesis in the rhesus monkey (Macaca mulatta) oviduct has been studied by stereomicroscopy, rotation photography, and serial section analysis. Two pathways for basal body production are described: acentriolar basal body formation (major pathway) where procentrioles are generated from a spherical aggregate of fibers; and centriolar basal body formation, where procentrioles are generated by the diplosomal centrioles. In both pathways, the first step in procentriole formation is the arrangement of a fibrous granule precursor into an annulus. A cartwheel structure, present within the lumen of the annulus, is composed of a central cylinder with a core, spoke components, and anchor filaments. Tubule formation consists of an initiation and a growth phase. The A tubule of each triplet set first forms within the wall material of the annulus in juxtaposition to a spoke of the cartwheel. After all nine A tubules are initiated, B and C tubules begin to form. The initiation of all three tubules occurs sequentially around the procentriole. Simultaneous with tubule initiation is a nonsequential growth of each tubule. The tubules lengthen and the procentriole is complete when it is about 200 mµ long. The procentriole increases in length and diameter during its maturation into a basal body. The addition of a basal foot, nine alar sheets, and a rootlet completes the maturation process. Fibrous granules are also closely associated with the formation of these basal body accessory structures.

scholarly journals THE RELATIONSHIP BETWEEN THE FINE STRUCTURE AND DIRECTION OF BEAT IN GILL CILIA OF A LAMELLIBRANCH MOLLUSC

1961 ◽  
Vol 11 (1) ◽  
pp. 179-205 ◽  
Author(s):  
I. R. Gibbons
Keyword(s):  
Fine Structure ◽  
Cell Surface ◽  
Basal Body ◽  
Transitional Region ◽  
Basal Bodies ◽  
Conical Structure ◽  
The Relationship ◽  
Basal Foot ◽  
Different Levels ◽  
Outer Fiber

This paper describes the fine structure and its relationship to the direction of beat in four types of cilia on the gill of the fresh-water mussel Anodonta cataracta. The cilia contain nine outer, nine secondary, and two central fibers, such as have been described previously in other material. Each outer fiber is a doublet with one subfiber bearing arms. One particular pair of outer fibers (numbers 5 and 6) are joined together by a bridge. The two central fibers are enclosed by a central sheath; also present in this region is a single, small mid-fiber. The different groups of fibers are connected together by radial links that extend from the outer to the secondary fibers, and from the secondary fibers to the central sheath. The basal body consists of a cylinder of nine triplet fibers. Projecting from it on one side is a dense conical structure called the basal foot. The cylinder of outer fibers continues from the basal body into the cilium, passing through a complex transitional region in which five distinct changes of structure occur at different levels. There are two sets of fibers associated with the basal bodies: a pair of striated rootlets that extends from each basal body down into the cell, and a system of fine tubular fibers that runs parallel to the cell surface. The relationship between fine structure and direction of beat is the same in all four types of cilia examined. The plane of beat is perpendicular to the plane of the central fibers, with the effective stroke toward the bridge between outer fibers 5 and 6, and toward the foot on the basal body.

scholarly journals Identification of contractile proteins in basal bodies of ciliated tracheal epithelial cells.

1980 ◽  
Vol 28 (11) ◽  
pp. 1189-1197 ◽  
Author(s):  
R E Gordon ◽  
B P Lane ◽  
F Miller
Keyword(s):  
Basal Body ◽  
Contractile Function ◽  
Ultrastructural Localization ◽  
Immunoperoxidase Technique ◽  
Basal Bodies ◽  
Tracheal Epithelial Cells ◽  
Tracheal Epithelial ◽  
Indirect Immunoperoxidase ◽  
Basal Foot ◽  
Basal Density

To determine the molecular composition of the components of basal bodies and the interbasal body apparatus of ciliated cells in rat tracheal epithelium, we used rabbit anti-actin, anti-alpha-actinin, anti-tropomyosin, and anti-myosin as primary antisera applied to the tissue in an indirect immunoperoxidase technique. The antisera was proven to be monospecific by elution of antibody after affinity chromatography. Sheep anti-rabbit immunoglobulin Fab fragments coupled to peroxidase were used for ultrastructural localization of the bound rabbit antibody. Antibodies against alpha-actinin were demonstrated around peripheral microtubules of cilia and linking these microtubules to central doublet and plasma membrane. Alpha-actinin was also shown in the basal foot processes. Anti-actin antibodies were associated with microtubules of the cilium and basal bodies, except in the region of the ciliary necklace. The antibodies directed against actin also had affinity for rootlets, basal foot processes, and communications between basal bodies and foot processes. Both anti-myosin and anti-tropomyosin antibodies were localized to part of the region of the constriction of the cilium, to the central basal density and the outer surfaces of basal body microtubules, and to the basal foot processes together with their communications to the basal body. The data suggest active contractile function of basal bodies.

scholarly journals The molecular dynamics of subdistal appendages in multi-ciliated cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Hyunchul Ryu ◽  
Haeryung Lee ◽  
Jiyeon Lee ◽  
Hyuna Noh ◽  
Miram Shin ◽  
...  
Keyword(s):  
Basal Body ◽  
Ependymal Cells ◽  
Basal Bodies ◽  
Ciliary Beating ◽  
Sam Domain ◽  
Adult Mice ◽  
Region I ◽  
Evolutionarily Conserved ◽  
Basal Foot

AbstractThe motile cilia of ependymal cells coordinate their beats to facilitate a forceful and directed flow of cerebrospinal fluid (CSF). Each cilium originates from a basal body with a basal foot protruding from one side. A uniform alignment of these basal feet is crucial for the coordination of ciliary beating. The process by which the basal foot originates from subdistal appendages of the basal body, however, is unresolved. Here, we show FGFR1 Oncogene Partner (FOP) is a useful marker for delineating the transformation of a circular, unpolarized subdistal appendage into a polarized structure with a basal foot. Ankyrin repeat and SAM domain-containing protein 1A (ANKS1A) interacts with FOP to assemble region I of the basal foot. Importantly, disruption of ANKS1A reduces the size of region I. This produces an unstable basal foot, which disrupts rotational polarity and the coordinated beating of cilia in young adult mice. ANKS1A deficiency also leads to severe degeneration of the basal foot in aged mice and the detachment of cilia from their basal bodies. This role of ANKS1A in the polarization of the basal foot is evolutionarily conserved in vertebrates. Thus, ANKS1A regulates FOP to build and maintain the polarity of subdistal appendages.

3-Hydroxy-3-methylglutaryl Coenzyme A Reductase from Spruce ( Picea abies). Properties and Regulation

1990 ◽  
Vol 45 (9-10) ◽  
pp. 973-979 ◽  
Author(s):  
Meinrad Boll ◽  
Angelika Kardinal
Keyword(s):  
Enzyme Activity ◽  
Picea Abies ◽  
Cell Suspension ◽  
Specific Activity ◽  
Reductase Activity ◽  
Cell Suspension Cultures ◽  
Callus Cultures ◽  
Suspension Cultures ◽  
Phase Cell ◽  
Ph Optimum

Abstract HM GCoA reductase was identified in seedlings, callus cultures, cell suspension cultures and in needles of spruce ( Picea abies) (L.) (Karst). Activity was found in both the 18 K pellet and in the 105 K pellet with different ratios between the two fractions from the various sources. The enzyme has a pH-optimum of 7.9 and an absolute requirement for NADPH . The presence of a thiol reagent such as dithiothreitol is required for activity. Km for HM G CoA is 20 -25 μM. Detergents have differential effects on the activity. In seedlings, enzyme activity was considerably higher in the hypocotyls than in the cotyledons. Enzyme activity was high in dark-grown and low in light-grown seedlings. When the light conditions were reversed, levels of activity adapted to the respective new conditions (increase or decline of specific activity). Aerobic incubations of seedlings, callus cultures or needles in medium containing a carbon source, resulted in a large (up to 20-fold) transient increase of HMGCoA reductase activity. Transfer of stationary phase cell suspension cultures into new medium caused a similarly large increase of activity. A number of carbohydrates induced the enzyme, glucose, fructose and sucrose being most effective. The increase of activity was prevented by cycloheximide. All changes of activity were much more pronounced in the 18 K pellet HMG CoA reductase

On the role of octopine dehydrogenase in cephalopod mantle muscle metabolism

1976 ◽  
Vol 54 (6) ◽  
pp. 871-878 ◽  
Author(s):  
Jeremy H. A. Fields ◽  
John Baldwin ◽  
Peter W. Hochachka
Keyword(s):  
Enzyme Activity ◽  
Muscle Activity ◽  
Specific Activity ◽  
Ph Dependence ◽  
Muscle Metabolism ◽  
Ph Optimum ◽  
Wet Weight ◽  
Octopine Dehydrogenase

Octopine dehydrogenases from the mantle muscle of the squid, Symplectoteuthis oualaniensis, and of the octopus, Octopus ornatus, were kinetically characterized and compared. In the squid, the specific activity of the enzyme was about 110 μmol product formed per minute per gram wet weight; in the octopus that value was over 600. Both enzymes show similar pH dependence; in the direction of octopine formation the pH optimum was about 6.5, whereas in the direction of octopine oxidation it was about 8.5. The affinities for NADH, arginine, and pyruvate were similar (Km values were about 0.04 mM, 7 mM, and 2 mM respectively). Increasing the concentration of either arginine or pyruvate increased the affinity for the cosubstrate (pyruvate or arginine), this mechanism being a means of regulating the enzyme activity in vivo. In the direction of octopine oxidation, the octopus enzyme showed a much higher affinity for octopine (Km = 0.8 mM) than did the squid enzyme (Km = 4.4 mM), suggesting that it may be better geared for reconverting octopine to arginine and pyruvate after anaerobic bursts of muscle activity.

Studies on isocitrate lyase isolated from Lupinus cotyledons

1979 ◽  
Vol 57 (9) ◽  
pp. 1131-1137 ◽  
Author(s):  
P. Vanni ◽  
M. T. Vincenzini ◽  
F. M. Nerozzi ◽  
S. P. Sinha
Keyword(s):  
Enzyme Activity ◽  
Solid State ◽  
Specific Activity ◽  
Isocitrate Lyase ◽  
Spectral Characteristics ◽  
Random Orientation ◽  
Ph Optimum ◽  
Helix Conformation ◽  
Α Helix ◽  
Fluorescence Peak

Isocitrate lyase (threo-Ds-isocitrate glyoxylate-lyase, EC 4.1.3.1) was isolated from cotyledons of Lupinus seedlings, purified 100-fold with respect to its initial specific activity and characterized (Km, pH optimum, Mg2+ requirement, sulfhydryl inhibitors, and synthase activity). The final purified preparation consisted of two homogeneous protein bands clearly separated by electrophoresis on polyacrylamide gel and chromatography on Sephadex G 200.Reducing agents are necessary for the maintenance of enzyme activity. The most effective reducing agent studied was 1,4-dithioerythreitol. The effect of several metabolites (oxalate, malonate, phosphoenolpyruvate, succinate, malate, tartrate, gluconate-6-phosphate, sorbose, sorbitol, and inositol) on the activity of purified preparations was tested. Oxalate proved to be the strongest inhibitor, seconded closely by phosphoenolpyruvate. The spectral characteristics of the purified enzyme are as follows: ultraviolet peak at 280 nm and fluorescence peak at 340 nm. The solid state infrared spectrum of the enzyme (lyophilized) showed that the enzyme was mostly in the α-helix conformation with very slight random orientation.

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