Characteristics of pollen diffusates and pollen wall cytochemistry in poplars

1980 ◽  
Vol 44 (1) ◽  
pp. 1-17
Author(s):  
A.E. Ashford ◽  
R.B. Knox
Keyword(s):  
Acid Phosphatase ◽  
Leucine Aminopeptidase ◽  
Populus Deltoides ◽  
Polyacrylamide Gel Electrophoresis ◽  
Structural Features ◽  
Populus Alba ◽  
Calcofluor White ◽  
Antigenic Activity ◽  
Extracellular Location ◽  
Isoenzyme Patterns

Pollen diffusates, known to be important in pollen-stigma interactions controlling interspecific incompatibility between Populus deltoides and Populus alba, have been partly characterized and shown to contain more than 20 protein bands by polyacrylamide gel electrophoresis, at least 4 of these being glycoproteins. Seven fractions had antigenic activity in rabbits and several enzyme activities were also present. Peroxidase and leucine aminopeptidase isoenzymes were detected in the diffusates, demonstrating the extracellular location of these 2 enzymes. Isoenzyme patterns of peroxidase, esterase and acid phosphatase were complex, with some bands common to both species. Localization of acid phosphatase in the intine and esterase in the exine was demonstrated after brief aldehyde fixation and low-temperature embedding in glycol methacrylate. The intine and exine sites were distinguished by their chemical and structural features. Calcofluor white M2R new proved to be an excellent stain for differentiating the intine. Aniline blue-positive material, probably beta-1,3-glucan, is present associated with the intine of many ungerminated as well as germinating grains: production of this material may be a response to damage.

The effect of 2,4-D and kinetin on the activity and isoenzyme pattern of various enzymes in cotyledon cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender)

1978 ◽  
Vol 56 (18) ◽  
pp. 2185-2195 ◽  
Author(s):  
Paul G. Arnison ◽  
W. G. Boll
Keyword(s):  
Acid Phosphatase ◽  
Phaseolus Vulgaris ◽  
Cell Suspension ◽  
Malate Dehydrogenase ◽  
Leucine Aminopeptidase ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Glutamate Oxaloacetate Transaminase ◽  
Cotyledon Cell ◽  
Isoenzyme Patterns

Changes in the activity and isoenzyme patterns of acid phosphatase, leucine aminopeptidase, glutamate-oxaloacetate transaminase, esterase, and malate and glutamate dehydrogenases were studied in cotyledon cell suspension cultures of Phaseolus vulgaris grown in the presence and absence of the growth regulators 2,4-dichlorophenoxyacetic acid and kinetin. With all enzymes studied, the pattern of isoenzymes and total enzymatic activity changed with the different phases of the culture cycle. In particular, the patterns of esterase, malate dehydrogenase, and glutamate dehydrogenase changed markedly with the inoculation of cells into fresh medium.The differences in isoenzyme patterns of cells grown with and without regulators were predominantly quantitative. However, certain minor isoenzymes of acid phosphatase, glutamate-oxaloacetate transaminase, esterase, and malate dehydrogenase were only detected in cultures grown in the presence of the regulators, while one isoenzyme of leucine aminopeptidase and two of esterase were unique to cells cultured in the absence of regulators.Three cathodic isoenzymes of acid phosphatase were released from wall material by 1 M NaCl. Such isoenzymes were also detected in the medium and in cytoplasmic extracts. Increase in the wall isoenzymes following inoculation into fresh medium was correlated with a decrease in anodic, cytoplasmic acid phosphatase.

Changes in isoenzyme patterns during imbibition and germination of lodgepole pine (Pinuscontorta var. latifolia)

1984 ◽  
Vol 14 (5) ◽  
pp. 743-746 ◽  
Author(s):  
J. A. Pitel ◽  
W. M. Cheliak ◽  
B. S. P. Wang
Keyword(s):  
Gel Electrophoresis ◽  
Leucine Aminopeptidase ◽  
Lodgepole Pine ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Glutamate Oxaloacetate Transaminase ◽  
Major Band ◽  
Tissue Specific ◽  
Isoenzyme Patterns

Isoenzymes of esterase (EST), glutamate-oxaloacetate transaminase (GOT), leucine aminopeptidase (LAP), and peroxidase (PER) were examined following various periods of imbibition and germination of lodgepole pine (Pinuscontorta var. latifolia Engelm.) seeds. The acidic and basic isoenzymes of embryos, and roots and shoots of germinating seedlings were analyzed by polyacrylamide gel electrophoresis. One major band of EST disappeared with imbibition, while some minor bands appeared and disappeared with imbibition and germination. Comparison of the roots and shoots after 7 and 14 days of germination showed several tissue-specific differences. The number of bands of GOT increased with imbibition and germination. The levels of activity of three isoenzymes differed between the root and shoot tissues. One band of LAP disappeared with imbibition. The levels of activity of two bands varied between the root and shoot tissues. The number of bands of PER increased dramatically following imbibition and germination. Also, many tissue-specific differences were observed between root and shoot tissues.

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 Biallelic neutrophil Na-antigen system is associated with a polymorphism on the phospho-inositol-linked Fc gamma receptor III (CD16)

Blood ◽  
1990 ◽  
Vol 75 (1) ◽  
pp. 213-217 ◽  
Author(s):  
TW Huizinga ◽  
M Kleijer ◽  
PA Tetteroo ◽  
D Roos ◽  
AE von dem Borne
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Specific Antigen ◽  
Phosphatidyl Inositol ◽  
Structural Features ◽  
Fc Gamma Receptor ◽  
Gamma Receptor ◽  
Sds Page ◽  
Antigen System ◽  
Genetically Determined

Neutrophils express two distinct types of receptor for the Fc region of IgG, FcRII and FcRIII, in amounts of 10,000 to 20,000 FcRII (40 Kd) and 100,000 to 200,000 FcRIII (50 to 80 Kd) per neutrophil. We showed that the FcRIII exhibits genetically determined heterogeneity, detectable by differences in electrophoretic mobility with sodium dodecyl sulfate (SDS) as well as by reaction with antibodies against the biallelic neutrophil-specific antigen system NA. FcRIII was precipitated with an FcRIII-specific monoclonal antibody (MoAb) from the neutrophils of 35 donors. NA1NA1 donors expressed an FcRIII with a molecular weight (mol wt) of 50 to 65 Kd, NA1NA2 donors expressed an FcRIII with a mol wt of 50 to 80 Kd, and NA2NA2 donors expressed an FcRIII with a mol wt of 65 to 80 Kd. Statistical analysis showed that the electrophoretic heterogeneity corresponds with the NA polymorphism (k = 1). Sequential immunoprecipitation with a MoAb against NA1 and a MoAb against anti- FcRIII, followed by SDS-polyacrylamide gel electrophoresis (PAGE), showed that NA1-FcRIII is distinct from NA2-FcRIII. Moreover, immunoprecipitation with a MoAb against NA1 yielded a protein of 50 to 65 Kd, and immunoprecipitation with human anti-NA2 sera or an MoAb against NA2 yielded a protein of 65 to 80 Kd. Preincubation of NA1NA2 neutrophils with F(ab')2 fragments of an MoAb against anti-NA1 reduced binding of IgG dimers to these cells with about 50%, whereas it completely prevented binding of the dimers to NA1NA1 neutrophils. Inhibition experiments with the MoAb against NA2 yielded the same results for NA1NA2 cells, whereas binding of IgG dimers to NA2NA2 cells was completely prevented. Thus, the products of both NA alleles bind IgG. Immunoprecipitation from the medium of neutrophils either stimulated with formyl- methionyl-leucyl-phenylalanine (FMLP) or treated with glycosyl-phosphatidyl-inositol-specific phospholipase C (GPI- PLC) showed that both the NA1-FcRIII and the NA2-FcRIII are released from the cell surface, indicating that both forms of FcRIII have some structural features in common. Deglycosylation of FcRIII from homozygous donors yielded material that showed several bands on SDS- PAGE. GPI-PLC treatment of neutrophils indicated that all of this material is phosphatidyl-inositol linked.

The isolation of surface array proteins from bacteria

1984 ◽  
Vol 62 (11) ◽  
pp. 1181-1189 ◽  
Author(s):  
S. F. Koval ◽  
R. G. E. Murray
Keyword(s):  
Molecular Weight ◽  
Protein Conformation ◽  
Gel Electrophoresis ◽  
Guanidine Hydrochloride ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Structural Features ◽  
Proteolytic Degradation ◽  
Low Concentrations ◽  
Single Polypeptide

The methods used for the isolation of regularly structured (RS) surface array proteins of a range of prokaryotes are described. Most RS proteins can be selectively solubilized from envelope preparations with low concentrations of urea or guanidine hydrochloride. Sodium dodecyl sulfate – polyacrylamide gel electrophoresis analysis of the protein extracts shows that most RS arrays are composed of a single polypeptide that may contain carbohydrate. The molecular weight of the proteins varies from 41 000 to 200 000. Possible reasons for the presence of more than one polypeptide in RS protein preparations are discussed, as well as the evidence for proteolytic degradation of some RS proteins during isolation. Structural features of the RS proteins are described and the importance of protein conformation to assembly of the arrays is indicated.

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.

Isoenzymes in cell cultures of bush bean (Phaseolus vulgaris cv. Contender): isoenzymatic differences between stock suspension cultures derived from a single seedling

1975 ◽  
Vol 53 (3) ◽  
pp. 261-271 ◽  
Author(s):  
Paul G. Arnison ◽  
W. G. Boll
Keyword(s):  
Acid Phosphatase ◽  
Phaseolus Vulgaris ◽  
Cell Cultures ◽  
Suspension Cultures ◽  
Peroxidase Isoenzymes ◽  
Bush Bean ◽  
Leucine Amino Peptidase ◽  
The Media ◽  
Isoenzyme Patterns ◽  
Stock Suspension

Electrophoretic analyses of isoenzyme patterns were performed with extracts of root, hypocotyl, and cotyledon suspension cultures derived from a single seedling. The enzymes studied included malate, glutamate, and glucose-6-phosphate dehydrogenases; peroxidase; polyphenol oxidase; esterase; acid phosphatase; and leucine amino peptidase. Peroxidase isoenzymes were also detected in the media. The isoenzymatic patterns of the three cultures were different for some enzymes, similar for others, and identical for the rest. The isoenzymatic patterns were recorded on a number of occasions over a period of 3 years and they remained relatively unchanged.

Fusion Expression on the Esat-6 and cfp-10 Genes of Mycobacterium bovis in Escherichia coli

2013 ◽  
Vol 421 ◽  
pp. 354-358
Author(s):  
Jia Ming Lin ◽  
Chun Fang Wang ◽  
Jia Ning Guan ◽  
Hong Xia Ma ◽  
Shuang Hou ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Mycobacterium Bovis ◽  
Expression Vector ◽  
Fusion Gene ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Antigenic Activity ◽  
Sds Page ◽  
Prokaryotic Expression Vector ◽  
Single Antigen

Based on the (Gly4Ser)3 linker, theesat-6andcfp-10gene were fused for raising the antigenicity of single antigen. The DNA fragments ofesat-6andcfp-10were fused by splicing by overlapping extension (SOE) polymerase chain reaction (PCR),and the fusion gene esat-6-cfp-10 were cloned into pMD18-T vector, and then we got the recombinant plasmid pMD-esat-6-cfp-10. pMD-esat-6-cfp-10 and pET28a (+) were digested byBamHI andEcoRI double enzymes. The purified mpb esat-6-cfp-10 fusion gene was subcloned into the expression vector pET28a (+),and the prokaryotic expression vector pET-esat-6-cfp-10 was constructed. Plasmid containing pET-esat-6-cfp-10 was transformed into competenceEscherichia coliBL21(DE3).The bacterium was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) and analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE),approximately 25 kDa exogenous protein was observed on the SDS-PAGE. The protein was analyzed by using Western-blotting. The results indicated that the protein was of antigenic activity ofMycobacterium bovis. These results could serve as a basis for further studies on the usefulness of the fusion gene and its expression product in the development of DNA vaccine; living carrier vaccine; subunit vaccine and diagnostic reagents against bovine tuberculosis.

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