Characterization of placental isoenzyme of alkaline phosphatase in human pregnancy serum

1969 ◽  
Vol 47 (2) ◽  
pp. 147-155 ◽  
Author(s):  
N. K. Ghosh ◽  
W. H. Fishman
Keyword(s):  
Alkaline Phosphatase ◽  
Serum Alkaline Phosphatase ◽  
Heat Stability ◽  
Biochemical Properties ◽  
Heat Inactivation ◽  
Total Serum ◽  
Placental Alkaline Phosphatase ◽  
Human Placental Alkaline Phosphatase ◽  
Alkaline Phosphatase Isoenzyme ◽  
In Pregnancy

Human placental alkaline phosphatase isoenzyme has been characterized in pregnancy serum by several biochemical criteria. The total serum alkaline phosphatase, its L-phenylalanine-sensitive moiety, heat inactivation, and the ratio of enzyme activity at pH 10.7 versus 9.8 (10.7/9.8 R) were measured during parturition and 59 weeks of pre- and post-natal periods. The extent of L-phenylalanine inhibition, heat stability, and 10.7/9.8 R of serum alkaline phosphatase progressively increased during gestation attaining maximum values during the delivery, after which they gradually declined. The electrophoretic behaviors of alkaline phosphatase isoenzymes of pregnancy sera were followed by starch- and Sephadex-gel electrophoreses. Alkaline phosphatase has been purified 300-fold from the placenta of the subject whose serum enzyme was investigated. The biochemical properties, including the electrophoretic behavior and neuraminidase sensitivity of heat-stable alkaline phosphastase in pregnancy sera at term, were comparable to those of purified placental alkaline phosphatase. The values for 10.7/9.8 R of the pregnancy sera were statistically different from those of sera from normal nonpregnant women. The results obtained in this study suggest that the enhanced level of pregnancy serum alkaline phosphatase is due to the enrichment of the circulation with an isoenzyme of placental origin.

Use of monoclonal antibodies to detect human placental alkaline phosphatase.

1983 ◽  
Vol 29 (1) ◽  
pp. 115-119 ◽  
Author(s):  
G De Groote ◽  
P De Waele ◽  
A Van de Voorde ◽  
M De Broe ◽  
W Fiers
Keyword(s):  
Monoclonal Antibody ◽  
Alkaline Phosphatase ◽  
Monoclonal Antibodies ◽  
Solid Phase ◽  
Enzymatic Reactions ◽  
Placental Alkaline Phosphatase ◽  
Human Placental Alkaline Phosphatase ◽  
Human Sera ◽  
Starch Gel ◽  
Alkaline Phosphatase Isoenzyme

Abstract Convenient, sensitive, and specific solid-phase immunoassays involving monoclonal antibody are described for the determination of human placental alkaline phosphatase (hPLAP). An endogenous enzyme immunoassay combined the specificity of the immunological and the enzymatic reactions. Alternatively, a solid-phase "sandwich" radioimmunoassay involving immobilized polyclonal rabbit anti-hPLAP in combination with iodinated monoclonal antibody provided some additional advantages. Both tests can be used to detect hPLAP from various sources, e.g., in human sera during pregnancy or as a tumor marker. The radioimmunoassay detected an increase in hPLAP at nine weeks of gestation. We discuss the use of monoclonal antibodies for the differentiation of different alkaline phosphatase isoenzyme types by electrophoresis on starch gel.

Hypophosphatasia (adult form): quantitation of serum alkaline phosphatase isoenzyme activity in a large kindred.

1980 ◽  
Vol 26 (7) ◽  
pp. 840-845 ◽  
Author(s):  
J L Millán ◽  
M P Whyte ◽  
L V Avioli ◽  
W H Fishman
Keyword(s):  
Alkaline Phosphatase ◽  
Serum Alkaline Phosphatase ◽  
Hepatic Metabolism ◽  
Total Activity ◽  
Heat Inactivation ◽  
Mean Values ◽  
Adult Form ◽  
Cellulose Acetate Membranes ◽  
Alkaline Phosphatase Isoenzymes ◽  
Alkaline Phosphatase Isoenzyme

Abstract We used heat inactivation, L-phenylalanine inhibition, and electrophoresis on polyacrylamide gel and cellulose acetate membranes--with and without use of specific antisera against the liver-bone, intestinal, and placental isoenzymes--to distinguish and quantitate the different alkaline phosphatase isoenzymes in sera from 23 adult members of a kindred affected by the adult form of hypophosphatasia. Nine subjects had values for total activity more than two standard deviations below the mean values for age- and sex-matched normal persons. Bone isoenzyme was diminished in all nine, whereas liver isoenzyme was subnormal in only four. Phosphoethanolamine and phosphoserine in the urine of eight hypophosphatasemic individuals correlated inversely with both total and liver alkaline phosphatase activity in their serum, but not with the activity of the bone isoenzyme. Total activity in the serum of adult kindred members correlated best with the circulating liver isoenzyme activity. The findings suggest that altered hepatic metabolism is responsible for the increased urinary excretion of phosphoethanolamine, and perhaps phosphoserine, in hypophosphatasia.

Atypical alkaline phosphatase isoenzyme in serum from a patient with Hodgkin's disease.

1984 ◽  
Vol 30 (5) ◽  
pp. 800-802 ◽  
Author(s):  
J P Beilby ◽  
P Garcia-Webb ◽  
C I Bhagat ◽  
A Prins
Keyword(s):  
Alkaline Phosphatase ◽  
Molecular Mass ◽  
Hodgkin's Disease ◽  
Serum Alkaline Phosphatase ◽  
Hodgkin’S Disease ◽  
Heat Inactivation ◽  
Relative Molecular Mass ◽  
Agarose Gel Electrophoresis ◽  
Post Translational Modification ◽  
Alkaline Phosphatase Isoenzyme

Abstract An alkaline phosphatase isoenzyme that did not move from the origin in agarose gel electrophoresis was detected in serum from a 51-year-old woman with Hodgkin's disease. Inhibitor and heat-inactivation studies of the patient's serum alkaline phosphatase showed properties resembling those of both liver and bone isoenzymes. No immunoglobulin or high-molecular-mass complexes with the alkaline phosphatase isoenzyme were detected. The relative molecular mass (Mr) of the atypical alkaline phosphatase isoenzyme was 182 000, that of the liver alkaline phosphatase isoenzyme control 170 000. Treatment of both of these isoenzymes with neuraminidase gave a product with an Mr of 140 000. We propose that a post-translational modification increased the carbohydrate content of the liver alkaline phosphatase isoenzyme, thus changing the charge characteristics of the enzyme and decreasing its electrophoretic mobility. We believe this to be the first report of a post-translational modification in a heat-sensitive isoenzyme of alkaline phosphatase.

scholarly journals Heat stability of human placental alkaline phosphatase

1965 ◽  
Vol 18 (3) ◽  
pp. 359-363 ◽  
Author(s):  
F. C. Neale ◽  
J. S. Clubb ◽  
D. Hotchkis ◽  
S. Posen
Keyword(s):  
Alkaline Phosphatase ◽  
Heat Stability ◽  
Placental Alkaline Phosphatase ◽  
Human Placental Alkaline Phosphatase

Hypophosphatasia (adult form): quantitation of serum alkaline phosphatase isoenzyme activity in a large kindred.

1980 ◽  
Vol 26 (7) ◽  
pp. 840-845
Author(s):  
J L Millán ◽  
M P Whyte ◽  
L V Avioli ◽  
W H Fishman
Keyword(s):  
Alkaline Phosphatase ◽  
Serum Alkaline Phosphatase ◽  
Hepatic Metabolism ◽  
Total Activity ◽  
Heat Inactivation ◽  
Mean Values ◽  
Adult Form ◽  
Cellulose Acetate Membranes ◽  
Alkaline Phosphatase Isoenzymes ◽  
Alkaline Phosphatase Isoenzyme

Abstract We used heat inactivation, L-phenylalanine inhibition, and electrophoresis on polyacrylamide gel and cellulose acetate membranes--with and without use of specific antisera against the liver-bone, intestinal, and placental isoenzymes--to distinguish and quantitate the different alkaline phosphatase isoenzymes in sera from 23 adult members of a kindred affected by the adult form of hypophosphatasia. Nine subjects had values for total activity more than two standard deviations below the mean values for age- and sex-matched normal persons. Bone isoenzyme was diminished in all nine, whereas liver isoenzyme was subnormal in only four. Phosphoethanolamine and phosphoserine in the urine of eight hypophosphatasemic individuals correlated inversely with both total and liver alkaline phosphatase activity in their serum, but not with the activity of the bone isoenzyme. Total activity in the serum of adult kindred members correlated best with the circulating liver isoenzyme activity. The findings suggest that altered hepatic metabolism is responsible for the increased urinary excretion of phosphoethanolamine, and perhaps phosphoserine, in hypophosphatasia.

Improved method for quantitative determination in serum of alkaline phosphatase of skeletal origin.

1981 ◽  
Vol 27 (12) ◽  
pp. 2002-2007 ◽  
Author(s):  
J R Farley ◽  
C H Chesnut ◽  
D J Baylink
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Serum Alkaline Phosphatase ◽  
Total Serum ◽  
Placental Alkaline Phosphatase ◽  
Serum Alkaline Phosphatase Activity ◽  
Total Body Calcium ◽  
Post Menopausal ◽  
Skeletal Alkaline Phosphatase

Abstract In this quantitative method for detection of skeletal alkaline phosphatase (EC 3.1.3.1) activity in human serum, intestinal and placental alkaline phosphatase activities are recognized by their susceptibility to inhibition by L-phenylalanine, and skeletal and hepatic alkaline phosphatases are distinguished by their different sensitivities to inactivation by heat. Alkaline phosphatase isoenzymes prepared from organ sources may behave differently from the corresponding isoenzymes in serum. Our procedure allows us to include organ-derived internal standards of skeletal, intestinal, and biliary alkaline phosphatase to minimize between-assay variation. In preliminary applications, we have found that (a) total serum alkaline phosphatase activity is extremely variable in post-menopausal osteoporotic subjects and is not a reliable index of skeletal alkaline phosphatase activity; (b) seven osteoporotic patients responding to therapy with sodium fluoride with increased bone formation showed increased skeletal alkaline phosphatase activity in their serum as compared with age-matched controls (p less than 0.005); and (c) 10 post-menopausal osteoporotic patients responding to therapy with stanozolol with increased total body calcium showed an increase in circulating skeletal alkaline phosphatase activity (p less than 0.001).

scholarly journals The Promoter of the Rat Gonadotropin-Releasing Hormone Receptor Gene Directs the Expression of the Human Placental Alkaline Phosphatase Reporter Gene in Gonadotrope Cells in the Anterior Pituitary Gland as well as in Multiple Extrapituitary Tissues

Endocrinology ◽  
2004 ◽  
Vol 145 (2) ◽  
pp. 983-993 ◽  
Author(s):  
Anne Granger ◽  
Valérie Ngô-Muller ◽  
Christian Bleux ◽  
Céline Guigon ◽  
Hanna Pincas ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Reporter Gene ◽  
Transgene Expression ◽  
Pituitary Cells ◽  
Placental Alkaline Phosphatase ◽  
R Gene ◽  
Glycoprotein Hormone ◽  
Steroidogenic Factor 1 ◽  
Functional Sites ◽  
Human Placental Alkaline Phosphatase

Abstract Previous studies dealing with the mechanisms underlying the tissue-specific and regulated expression of the GnRH receptor (GnRH-R) gene led us to define several cis-acting regulatory sequences in the rat GnRH-R gene promoter. These include functional sites for steroidogenic factor 1, activator protein 1, and motifs related to GATA and LIM homeodomain response elements as demonstrated primarily in transient transfection assays in mouse gonadotrope-derived cell lines. To understand these mechanisms in more depth, we generated transgenic mice bearing the 3.3-kb rat GnRH-R promoter linked to the human placental alkaline phosphatase reporter gene. Here we show that the rat GnRH-R promoter drives the expression of the reporter gene in pituitary cells expressing the LHβ and/or FSHβ subunit but not in TSHβ- or GH-positive cells. Furthermore, the spatial and temporal pattern of the transgene expression during the development of the pituitary was compatible with that characterizing the emergence of the gonadotrope lineage. In particular, transgene expression is colocalized with the expression of the glycoprotein hormone α-subunit at embryonic day 13.5 and with that of steroidogenic factor 1 at later stages of pituitary development. Transgene expression was also found in specific brain areas, such as the lateral septum and the hippocampus. A single promoter is thus capable of directing transcription in highly diverse tissues, raising the question of the different combinations of transcription factors that lead to such a multiple, but nevertheless cell-specific, expressions of the GnRH-R gene.

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