scholarly journals Structural and kinetic properties of a novel purple acid phosphatase from phosphate-starved tomato (Lycopersicon esculentum) cell cultures

2004 ◽  
Vol 377 (2) ◽  
pp. 419-428 ◽  
Author(s):  
Gale G. BOZZO ◽  
Kashchandra G. RAGHOTHAMA ◽  
William C. PLAXTON
Keyword(s):  
Acid Phosphatase ◽  
Lycopersicon Esculentum ◽  
Peptide Mapping ◽  
Periodic Acid ◽  
Purple Acid Phosphatase ◽  
Kinetic Properties ◽  
Suspension Cells ◽  
Periodic Acid Schiff ◽  
Α Subunit ◽  
Α And Β Subunits

An intracellular acid phosphatase (IAP) from Pi-starved (−Pi) tomato (Lycopersicon esculentum) suspension cells has been purified to homogeneity. IAP is a purple acid phosphatase (PAP), as the purified protein was violet in colour (λmax=546 nm) and was insensitive to l-tartrate. PAGE, periodic acid–Schiff staining and peptide mapping demonstrated that the enzyme exists as a 142 kDa heterodimer composed of an equivalent ratio of glycosylated and structurally dissimilar 63 (α-subunit) and 57 kDa (β-subunit) polypeptides. However, the nine N-terminal amino acids of the α- and β-subunits were identical, exhibiting similarity to the deduced N-terminal portions of several putative plant PAPs. Quantification of immunoblots probed with rabbit anti-(tomato acid phosphatase) immune serum revealed that the 4-fold increase in IAP activity due to Pi-deprivation was correlated with similar increases in the amount of antigenic IAP α- and β-subunits. IAP displayed optimal activity at pH 5.1, was activated 150% by 10 mM Mg2+, but was potently inhibited by Zn2+, Cu2+, Fe3+, molybdate, vanadate, fluoride and Pi. Although IAP demonstrated broad substrate selectivity, its specificity constant (Vmax/Km) with phosphoenolpyruvate was >250% greater than that obtained with any other substrate. IAP exhibited significant peroxidase activity, which was optimal at pH 9.0 and insensitive to Mg2+ or molybdate. This IAP is proposed to scavenge Pi from intracellular phosphate esters in −Pi tomato. A possible secondary IAP role in the metabolism of reactive oxygen species is discussed. IAP properties are compared with those of two extracellular PAP isoenzymes that are secreted into the medium of −Pi tomato cells [Bozzo, Raghothama and Plaxton (2002) Eur. J. Biochem. 269, 6278–6286].

Prefertilization ovule development in Capsella: the dyad, tetrad, developing megaspore, and two-nucleate gametophyte

1986 ◽  
Vol 64 (4) ◽  
pp. 875-884 ◽  
Author(s):  
Patricia Schulz ◽  
William A. Jensen
Keyword(s):  
Cell Wall ◽  
Electron Microscope ◽  
Acid Phosphatase ◽  
De Novo ◽  
Periodic Acid ◽  
Aniline Blue ◽  
Ovule Development ◽  
Periodic Acid Schiff ◽  
Fluorescent Material ◽  
Autophagic Vacuoles

Ovules of Capsella bursa-pastoris at the dyad and tetrad stages of meiosis and at the megaspore and two-nucleate stages of the gametophyte were studied with the electron microscope. The cells of the dyad and tetrad are separated by aniline blue fluorescent cross walls and receive all types of organelles and autophagic vacuoles that were present in the meiocyte. Autophagic vacuoles enclose ribosomes and organelles and show reaction product for acid phosphatase. Autophagic vacuoles and some plastids are absorbed into the enlarging vacuoles of the growing megaspore. Other plastids appear to survive meiosis and there is no evidence for their de novo origin. Some mitochondria appear to degenerate in the enlarging megaspore but others look healthy and there is no evidence for the de novo origin of mitochondria. The nucleolus of the developing megaspore becomes very large and the cytoplasm is extremely dense with ribosomes. The cell wall is thickened by an electron-translucent, periodic acid – Schiff negative, aniline blue fluorescent material and contains plasmodesmata that link the megaspore with the nucellus. The plasmalemma of the growing megaspore produces microvilluslike extensions into this wall that disappear with the formation of the two-nucleate gametophyte. Plasmodesmata disappear from the cell wall at the four-nucleate stage.

The structure and associations of the double S layer on the cell wall of Aquaspirillum sinuosum

1990 ◽  
Vol 36 (5) ◽  
pp. 327-335 ◽  
Author(s):  
Stephen H. Smith ◽  
Robert G. E. Murray
Keyword(s):  
Cell Wall ◽  
Outer Membrane ◽  
Outer Layer ◽  
Peptide Mapping ◽  
Periodic Acid ◽  
Minor Component ◽  
Membrane Resistance ◽  
Surface Layers ◽  
Periodic Acid Schiff ◽  
Bacterial Surface

Aquaspirillum sinuosum cell walls bear two paracrystalline, proteinaceous surface layers (S layers). Each shows a different symmetry: the inner layer is closely apposed to the outer membrane and is a tetragonal array (90° axes; 5-nm units; repeat frequency 8 nm); the outer layer is a hexagonal array on the external surface (14-nm units; repeat frequency 18 nm) and, although the units have a six-pointed stellate form, the linkage between units is not resolved. The outer layer consists of a major 130-kDa protein and a 180-kDa minor component; these co-extract, co-assemble, and are inseparable by hydroxylapatite chromatography or by recrystallization. The solubilizing effects of reagents suggest stabilization by hydrogen bonding and Ca2+. The two outer layer proteins are serologically related and show partial identity by peptide mapping. Periodic acid – Schiff staining of the 180-kDa band suggests that this may be a glycosylated form of the 130-kDa component. The inner layer components form a doublet of 75- and 80-kDa polypeptides with extreme resistance to extraction. Close apposition to the outer membrane, resistance to chaotropes, aqueous insolubility, and behaviour in charge-shift electrophoresis suggest hydrophobic interaction between subunits and an integral association with the outer membrane. Key words: bacterial surface, cell wall, surface layers, cell-wall proteins, cell-wall assembly.

The blood leukocytes of Bufo marinus: a light, phase-contrast, and histochemical study

1987 ◽  
Vol 65 (6) ◽  
pp. 1445-1453 ◽  
Author(s):  
M. Samuel Cannon ◽  
H. W. Sampson ◽  
E. D. Kapes
Keyword(s):  
Acid Phosphatase ◽  
Phase Contrast ◽  
Periodic Acid ◽  
Hydrolytic Enzymes ◽  
Neutral Red ◽  
Bufo Marinus ◽  
Oxidative Enzymes ◽  
Blood Leukocytes ◽  
Periodic Acid Schiff ◽  
Aerobic And Anaerobic

Blood leukocytes of Bufo marinus were studied by light and phase-contrast microscopy and histochemical techniques for the localization of glycogen, lipids, several basic proteins, and a number of hydrolytic and oxidative enzymes. The hydrolytic enzymes occurred in varying amounts in neutrophils, eosinophils, lymphocytes, and monocytes; neutrophils were the only leukocytes to demonstrate alkaline phosphatase activity, while β-glucuronidase was only seen in lymphocytes, and aryl-sulfatase was not observed in any leukocytes. Periodic acid – Schiff (PAS) positive granules also occurred in varying amounts in leukocytes. Slight lipid activity was only seen in neutrophils, while arginine, and (or) lysine, and tyrosine reactivity was only observed in eosinophils. The appearance and histochemical reactivity of acid phosphatase granules in neutrophils corresponded closely with the appearance and number of specific neutrophilic granules seen in Wright–Giemsa preparations and with the PAS-positive granules. Small lymphocytes were myeloperoxidase (peroxidase) negative; β-glucuronidase, acid phosphatase, and PAS-positive granules corresponded to neutral red granules seen in supravital films. The oxidative enzymes also occurred in differing amounts in leukocytes, but strongly suggested that the leukocytes of Bufo marinus are capable of some degree of aerobic and anaerobic metabolism.

Acid phosphatase localization in PAS-bodies of Gonyaulax

1981 ◽  
Vol 51 (1) ◽  
pp. 15-23
Author(s):  
R.E. Schmitter ◽  
A.J. Jurkiewicz
Keyword(s):  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Gonyaulax Polyedra ◽  
Marine Dinoflagellate ◽  
Pas Staining ◽  
First Time

Periodic acid-Schiff staining, acid phosphatase localization, and yellow autofluorescence have been correlated with the PAS-body of Gonyaulax polyedra for the first time. PAS- staining and acid phosphatase activity are both correlated with the PAS-body of Gonyaulax tamarensis. These results that the PAS-body of these marine dinoflagellate algae functions in subcellular digestion.

scholarly journals Biochemical and Molecular Characterization of AtPAP26, a Vacuolar Purple Acid Phosphatase Up-Regulated in Phosphate-Deprived Arabidopsis Suspension Cells and Seedlings

2006 ◽  
Vol 142 (3) ◽  
pp. 1282-1293 ◽  
Author(s):  
Vasko Veljanovski ◽  
Barbara Vanderbeld ◽  
Vicki L. Knowles ◽  
Wayne A. Snedden ◽  
William C. Plaxton
Keyword(s):  
Acid Phosphatase ◽  
Molecular Characterization ◽  
Purple Acid Phosphatase ◽  
Suspension Cells

The blood leukocytes of Bufo alvarius: a light, phase-contrast, and histochemical study

1979 ◽  
Vol 57 (2) ◽  
pp. 314-322 ◽  
Author(s):  
M. Samuel Cannon ◽  
A. M. Cannon
Keyword(s):  
Acid Phosphatase ◽  
Phase Contrast ◽  
Periodic Acid ◽  
Hydrolytic Enzymes ◽  
Neutral Red ◽  
Nonspecific Esterase ◽  
Alkaline Phosphatases ◽  
Blood Leukocytes ◽  
Periodic Acid Schiff ◽  
Aryl Sulfatase

Blood leukocytes of Bufo alvarius were studied by light and phase-contrast microscopy and histochemical techniques for the localization of glycogen and several hydrolytic enzymes, i.e., acid and alkaline phosphatases, nonspecific esterase, beta-glucuronidase, aryl-sulfatase, and myeloperoxidase (peroxidase). Neutrophils were the only leukocytes to demonstrate alkaline phosphatase activity, while beta-glucuronidase and aryl-sulfatase were not observed in any leukocytes. Periodic acid – Schiff (PAS) positive granules and granules containing hydrolytic enzymes occurred in varying amounts in leukocytes. In eosinophils, most glycogen was associated with smaller granules, while the larger refractile granules were PAS negative. Small lymphocytes were myeloperoxidase (peroxidase) negative. The present study agrees with previous investigations in mammals which indicate that specific granules in granulocytes may be PAS positive as well as contain one or more hydrolytic enzymes. In small lymphocytes of B. alvarius, PAS positive and acid phosphatase positive granules correspond to neutral red granules seen in supravital films. Furthermore, the appearance and histochemical reactivity of acid phosphatase granules in mature neutrophils, metamyelocytes, and late myelocytes correspond closely with the appearance and number of specific neutrophilic granules seen in Wright–Giemsa preparations and with PAS positive granules.

66 HISTIOTROPHIC NUTRITION AND IRON TRANSFER ACROSS THE MATERNO-FETAL INTERFACE IN NUCLEAR TRANSFER-DERIVED SOMATIC CATTLE CLONES

2006 ◽  
Vol 18 (2) ◽  
pp. 141 ◽  
Author(s):  
F. Pereira ◽  
F. Meirelles ◽  
F. Braga ◽  
J. Visintin ◽  
R. Rumpf ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Periodic Acid ◽  
Cell Debris ◽  
Iron Transfer ◽  
Periodic Acid Schiff ◽  
Endometrial Stroma ◽  
Transporter Protein ◽  
Uterine Gland ◽  
Uterine Glands ◽  
Phosphatase Enzyme

The histiotrophic nutrition by the endometrial glands and the materno-fetal interface in the cloned cattle placenta were analyzed in order to investigate the iron transfer. Placentomes and intercaruncular region samples were recovered at term Caesarean delivery from 14 cloned cattle and 10 controls, fixed in 4% paraformaldehyde and 10% formaldehyde in PBS, processed and stained for light microscopy (hematoxylin-eosin, picrosirius, and Masson's trichrome), histochemistry [Perls, acid phosphatase and periodic acid Schiff (PAS) reactions], and immunohistochemistry (with rabbit anti-pig uteroferrin antibody because the uteroferrin is an iron transporter protein). In the controls we verified blood extravasations in the materno-fetal interface between the uterine and the trophoblast epithelium characterized by hemophagous areas with consequent erythrophagocytosis by the adjacent trophoblast. This content presented extravasated erythrocytes, plasm, cell debris, and cells in a probable apoptotic process. The Perls histochemical reactions that exposed the ferric iron in the placentomes were positive, as was the uteroferrin immunohistochemistry in the trophoblast cytoplasm and in other deep points in the placentomes. The histochemical reactions, demonstrating the acid phosphatase enzyme that detects the phagocytic activity, were positive in the mesenchyme and trophoblast, with a weak stain in an endometrial stroma. In the top of fetal villi, mainly in the binucleate cells, we visualized accumulations of PAS-positive secretions, indicating the presence of mucoid material. The uterine gland epithelium was columnar-type and in the gland lumina there were cell debris and PAS-positive mucoid secretions. We confirmed the reactivity of the uterine glands to the acid phosphatase enzyme and to the Perls reaction in the epithelium and in the gland lumina. The uteroferrin immunohistochemistry showed a strong stain in the cytoplasm of the endometrial glands cells and in the lumina. In the NT bovine placentae, the blood extravasations between uterine and trophoblast epithelium were aberrant. There was also the remodeling of the maternal connective tissue (endometrial stroma) in this area. We also demonstrated phagocytic uptake of uteroferrin by the trophoblast, although the histochemical and immunohistochemical reactions were weak in the trophoblast of the placentomes and in the endometrial glands of the intercaruncular region, when compared with the controls. The results obtained by the histochemistry and immunohistochemistry indicated that these sites of transfer substances from mother to fetus are very important in providing adequate nutrition to the fetus, key to a successful pregnancy in NT bovines. This work was funded by FAPESP, Brazil.

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