The enzymic deacylation of esterified mono- and di-saccharides. I. The isolation and purification of an esterase from wheat germ lipase

1969 ◽  
Vol 47 (2) ◽  
pp. 135-142 ◽  
Author(s):  
A. L. Fink ◽  
G. W. Hay
Keyword(s):  
Acid Phosphatase ◽  
Wheat Germ ◽  
Polyacrylamide Gel Electrophoresis ◽  
Disc Electrophoresis ◽  
Isolation And Purification ◽  
Electrophoretic Behavior ◽  
Commercial Enzymes ◽  
Cursory Examination ◽  
Different Sources ◽  
Molecular Sieving

Whereas wheat germ lipase was known to contain acid phosphatase (EC 3.1.3.2), esterase (EC 3.1.1.1), and lipase (EC 3.1.1.3) activity, a cursory examination of some commercial enzymes has demonstrated also the presence of phosphodiesterase (EC 3.1.4.1), α- and β-glucosidase (EC 3.2.1.20 and EC 3.2.1.21), α- and β-galactosidase (EC 3.2.1.22 and EC 3.2.1.23), α-mannosidase (EC 3.2.1.24), and β-xylosidase (EC 3.2.1.37) activities. Disc electrophoresis revealed a minimum of 12 electrophoretically distinct protein fractions. Different sources and different batches of enzyme had similar disc electrophoretic behavior and activities. Fractionation of the crude preparation on carboxymethylcellulose or Sephadex, and electrophoresis on Sephadex, failed to separate α-glucosidase and esterase activities. The inhibition of α- and β-glucosidase could be effected by D-glucono-1,5-lactone, but not by metal salts or Tris.Preparative polyacrylamide-gel electrophoresis of wheat germ lipase gave a large number of active zones including several esterases, acid phosphatase, and α- and β-glucosidase fractions. Evidence was established for the nonidentity of two neighboring esterase zones. One was found to be free of α- and β-glucosidase activity. Sulfhydryl reagents inhibited the esterases, whereas acetone activated some and inactivated others. The esterases were most stable at neutral pH and were extensively inactivated on lyophilization, dialysis, and desalting with molecular-sieving reagents. Some properties of the purified esterase were examined.

scholarly journals Specific glycoprotein changes during development of the chick neural retina.

1978 ◽  
Vol 79 (1) ◽  
pp. 132-137 ◽  
Author(s):  
G Mintz ◽  
L Glaser
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Neural Retina ◽  
Cell Type ◽  
Retinal Antigens ◽  
Qualitative Changes

After separation of whole proteins of chick neural retina by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS), a number of glycoproteins can be detected by staining the gels with 125I-labeled wheat germ agglutinin (WGA) and other lectins. The glycoprotein patterns show both quantitative and qualitative changes between days 7 and 13 of development. Some of these glycoproteins can be separated by chromatography on columns of insolubilized lectins. These observations suggest that purification of some of these glycoproteins identified by staining with radioactive lectins would yield retinal antigens which may be specific for developmental stage and cell type.

LA PHOSPHATASE ACIDE DU GERME DE BLÉ: ANALYSE CHROMATOGRAPHIQUE

1965 ◽  
Vol 43 (12) ◽  
pp. 1899-1905 ◽  
Author(s):  
Jacques Brouillard ◽  
Ludovic Ouellet
Keyword(s):  
Acid Phosphatase ◽  
Chromatographic Analysis ◽  
Wheat Germ ◽  
Molecular Forms ◽  
Phosphatase Acide ◽  
Visible Spectra

Chromatographic analysis of acid phosphatase of wheat germ shows that this enzyme exists as four different, active, and easily separable molecular forms. The four fractions also hydrolyze esterase substrates. All four isozymes are yellow, and the visible spectra of three of them are given. Following a previous observation, we confirmed the presence of iron in the enzyme, presumably as Fe+++.

scholarly journals Assessment of Acid Phosphatase Production by In vitro Cultures of Atropa acuminata

2016 ◽  
Vol 26 (1) ◽  
pp. 15-23
Author(s):  
Saima Khan ◽  
Meenu Katoch ◽  
Sharada Mallubhotla ◽  
Suphla Gupta ◽  
Manju Sambyal ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Growth Period ◽  
Callus Cultures ◽  
Shoot Cultures ◽  
Crude Enzyme Extract ◽  
Atropa Acuminata

The potential of various culture lines of Atropa acuminata were investigated for resourcing acid phosphatase (ACP) (3.1.3.2). Crude enzyme extract comprised of a mixture of four isoforms, distinguishable by polyacrylamide gel electrophoresis (PAGE) with molecular weight ranging from 39 to 215 kDa. In vitro regenerated proliferative shoots, callus and roots showed higher specific activity (2.49, 3.41, 2.91 U/mg protein, respectively) as compared to in vivo grown plants (0.71 U/mg protein). ACP activity in root cultures increased progressively up to 4.6 U/mg during the entire growth period (2 ? 24 weeks), whereas in case of shoot cultures, the specific activity escalated to 2.49 U/mg at 8 weeks, which then declined subsequently (1.95 U/mg). Similarly, callus cultures initially showed a higher phosphohydrolytic activity (3.41 U/mg protein) until 8 weeks by which period, it decreased with the passage of growth period. The present studies reveal an alternate system for resourcing of ACP from Atropa acuminata.Plant Tissue Cult. & Biotech. 26(1): 15-23, 2016 (June)

In vitro translation of human prostatic acid phosphatase mRNA and processing of the translation products by microsomal membranes and endoglycosidase H

1987 ◽  
Vol 65 (10) ◽  
pp. 921-924 ◽  
Author(s):  
Gilles Paradis ◽  
Jean Y. Dubé ◽  
Pierre Chapdelaine ◽  
Roland R. Tremblay
Keyword(s):  
Acid Phosphatase ◽  
Molecular Mass ◽  
Polyacrylamide Gel Electrophoresis ◽  
Prostatic Acid Phosphatase ◽  
In Vitro Translation ◽  
Acid Phosphatases ◽  
Major Band ◽  
Microsomal Membranes ◽  
Endoglycosidase H

Poly(A)+ RNA was isolated from human prostatic tissue and translated in vitro in a rabbit reticulocyte lysate translation assay. Acid phosphatase labeled with [35S]methionine was immunoprecipitated with an antibody against seminal plasma acid phosphatase. Two-dimensional polyacrylamide gel electrophoresis of the immunoprecipitate, followed by fluorography, revealed the presence of two spots (one major and one minor), both having a molecular mass of 43 kilodaltons (kDa) and an isoelectric point higher than mature acid phosphatase. Addition of canine pancreatic membranes to the translation assay resulted in the formation of four immunoprecipitable spots with molecular masses ranging from 43 to 49 kDa on one-dimensional gels. These spots probably represent acid phosphatases containing one to four core sugar groups, since after the addition of endoglycosidase H the molecular mass heterogeneity was abolished and we observed only one major band with a molecular mass (41 kDa) slightly lower than the ones of the primary translation product. These results suggest that human prostatic acid phosphatases are synthesized as two 43-kDa preproteins, which are further processed to 41-kDa proteins by removal of their signal peptide. Heterogeneity of the native protein arises mostly from glycosylation at four sites and not from differences in the amino acid sequence of the various forms.

scholarly journals Wheat Germ Acid Phosphatase Activity in High Percentages of Organic Solvents

2020 ◽  
Vol 34 (S1) ◽  
pp. 1-1
Author(s):  
Veronica R. Moorman ◽  
Evan D. Kindl ◽  
Fiona N. Summers
Keyword(s):  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Organic Solvents ◽  
Acid Phosphatase Activity ◽  
Wheat Germ

scholarly journals Trichomonas vaginalis Lipophosphoglycan Mutants Have Reduced Adherence and Cytotoxicity to Human Ectocervical Cells

2005 ◽  
Vol 4 (11) ◽  
pp. 1951-1958 ◽  
Author(s):  
Felix D. Bastida-Corcuera ◽  
Cheryl Y. Okumura ◽  
Angie Colocoussi ◽  
Patricia J. Johnson
Keyword(s):  
Wheat Germ Agglutinin ◽  
Parent Strain ◽  
Trichomonas Vaginalis ◽  
Wheat Germ ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Host Cells ◽  
Chemical Mutagenesis ◽  
Reduced Adherence

ABSTRACT The extracellular human pathogen Trichomonas vaginalis is covered by a dense glycocalyx thought to play a role in host-parasite interactions. The main component of the glycocalyx is lipophosphoglycan (LPG), a polysaccharide anchored in the plasma membrane by inositol phosphoceramide. To study the role of LPG in trichomonads, we produced T. vaginalis LPG mutants by chemical mutagenesis and lectin selection and characterized them using morphological, biochemical, and functional assays. Two independently selected LPG mutants, with growth rates comparable to that of the wild-type (parent) strain, lost the ability to bind the lectins Ricinnus comunis agglutinin I (RCA120) and wheat germ agglutinin, indicating alterations in surface galactose and glucosamine residues. LPG isolated from mutants migrated faster than parent strain LPG on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting the mutants had shorter LPG molecules. Dionex high-performance anion exchange chromatography with pulsed amperometric detection analyses revealed galactosamine, glucosamine, galactose, glucose, mannose/xylose, and rhamnose as the main monosaccharides of T. vaginalis parent strain LPG. LPG from both mutants showed a reduction of galactose and glucosamine, corresponding with the reduced size of their LPG molecules and inability to bind the lectins RCA120 and wheat germ agglutinin. Mutant parasites were defective in attachment to plastic, a characteristic associated with avirulent strains of T. vaginalis. Moreover, the mutants were less adherent and less cytotoxic to human vaginal ectocervical cells in vitro than the parental strain. Finally, while parent strain LPG could inhibit the attachment of parent strain parasites to vaginal cells, LPG from either mutant could not inhibit attachment. These combined results demonstrate that T. vaginalis adherence to host cells is LPG mediated and that an altered LPG leads to reduced adherence and cytotoxicity of this parasite.

Size analysis of uncoupling protein and its precursor from brown adipose tissue of different species

1985 ◽  
Vol 63 (9) ◽  
pp. 988-991 ◽  
Author(s):  
Karl B. Freeman ◽  
Karen Meyrick ◽  
Hasmukh V. Patel ◽  
Robert G. Ridley
Keyword(s):  
Adipose Tissue ◽  
Brown Adipose Tissue ◽  
Uncoupling Protein ◽  
Mammalian Species ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Apparent Size ◽  
Size Analysis ◽  
Brown Adipose ◽  
Different Sources

The apparent size of the monomeric form of uncoupling protein from brown adipose tissue of several mammalian species was compared by sodium dodecyl sulfate – polyacrylamide gel electrophoresis. Including earlier results the apparent molecular mass of the protein from rat was about 32 000 daltons and varied about 1500–2000 daltons from the different sources with the size increasing in the order human < rat ≤ mouse ≤ hamster ≤ rabbit. The size of newly synthesized uncoupling protein was also found to vary among species. However in the two cases examined, rat and rabbit, the precursor and its respective mature monomeric protein had the same apparent size as shown by coelectrophoresis.

Acid phosphatase isoforms in dry seeds and during seedling development in barley (Hordeum vulgare)

1996 ◽  
Vol 74 (5) ◽  
pp. 653-658
Author(s):  
S. Pasqualini ◽  
P. Batini ◽  
L. Ederli ◽  
F. Panara ◽  
M. Antonielli
Keyword(s):  
Cell Wall ◽  
Acid Phosphatase ◽  
Hordeum Vulgare ◽  
Developmental Stages ◽  
Soluble Fraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Seedling Development ◽  
Rapid Decrease ◽  
Hordeum Vulgare L ◽  
Electrophoretic Patterns

The acid phosphatase activity in the soluble, membrane, and cell wall fractions from Hordeum vulgare in dry seeds and during seedling development was investigated. The acid phosphatase activities were also assayed in barley roots and coleoptiles at different developmental stages. Electrophoretic patterns of multiple acid phosphatases in seeds, endosperms and embryos, and growing roots and coleoptiles are shown. The enzyme activity shows a rapid decrease in both roots and coleoptiles during growth. Using nondenaturing polyacrylamide gel electrophoresis, multiple acid phosphatase forms were found in all the organs examined. However, no qualitative differences in the location of bands were observed between root and coleoptile extract at various stages of development. The coleoptile cell wall fraction showed an acid phosphatase form characterized by a very low electrophoretic mobility that was not found in the soluble fraction. Keywords: barley, Hordeum vulgare L., acid phosphatase, isoforms, seedlings growth.

EFFECTS OF 5α-ANDROSTANE-3β,17β-DIOL AND 5β-DIHYDROTESTOSTERONE ON ACID PHOSPHATASE ACTIVITY IN THE PROSTATE GLAND OF THE CASTRATED ADULT RAT

1978 ◽  
Vol 79 (1) ◽  
pp. 9-16 ◽  
Author(s):  
M. P. TENNISWOOD ◽  
PAMELA P. ABRAHAMS ◽  
C. E. BIRD ◽  
A. F. CLARK
Keyword(s):  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Gel Electrophoresis ◽  
Acid Phosphatase Activity ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Prostate Gland ◽  
Intraperitoneal Administration ◽  
Adult Rat ◽  
Secretory Acid Phosphatase

Polyacrylamide gel electrophoresis of filtrates from adult rat prostatic tissue showed two bands of acid phosphatase activity. These corresponded to the lysosomal and secretory acid phosphatases. After castration the secretory acid phosphatase disappeared. The specific activity of the enzyme increased from the time of castration to a maximum on day 7 before declining steadily, while the percentage inhibition by tartrate of acid phosphatase increased from control levels to a maximum on day 7 and then decreased to a new steady state by day 15. When 5α-androstane-3β,17β-diol was administered i.p. at a dose of 2 mg/day, starting immediately after castration, the secretory acid phosphatase was retained but the percentage inhibition and the specific activity were both raised above control levels. When this steroid was administered daily starting 7 days after castration the secretory acid phosphatase band on the gels returned more rapidly than with the classical androgens, but the percentage inhibition and specific activity were once again raised. Intraperitoneal administration of 5β-dihydrotestosterone, at a dose of 2 mg/day, did not maintain the secretory acid phosphatase activity which disappeared by day 5. However, the specific activity of acid phosphatase and the percentage inhibition by tartrate were both raised throughout the experiment. If this steroid was given 7 days after castration, the percentage inhibition by tartrate did not respond and fell to the level seen in castrated rats. The specific activity, however, remained significantly above the level found in castrated control rats.

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