Graphical techniques for kinetic data analyses of alkaline phosphatase

1980 ◽  
Vol 52 (11) ◽  
pp. 1730-1738 ◽  
Author(s):  
Jack W. Frazer ◽  
Hal R. Brand
Keyword(s):  
Alkaline Phosphatase ◽  
Kinetic Data ◽  
Data Analyses

SOME PROPERTIES OF AN ALKALINE PHOSPHATASE FROM RAT ADIPOSE TISSUE

1964 ◽  
Vol 42 (10) ◽  
pp. 1445-1457 ◽  
Author(s):  
Donald P. Wallach ◽  
Howard Ko
Keyword(s):  
Alkaline Phosphatase ◽  
Adipose Tissue ◽  
Kinetic Data ◽  
Aqueous Extracts ◽  
Partial Inhibition ◽  
Metallic Cation ◽  
Inhibition Kinetic ◽  
Salient Features ◽  
Rat Adipose Tissue ◽  
Ethylene Diaminetetraacetic Acid

An alkaline phosphatase was partially purified from aqueous extracts of rat adipose tissue, and some characteristics of the enzyme delineated. Among salient features, the enzyme is strongly inhibited by cysteine and ethylene diaminetetraacetic acid (EDTA) and types of inhibition were determined. Cysteine induced "complete exclusive" inhibition, while EDTA caused "partial" inhibition. Kinetic data on the inhibition by EDTA are consistent with a mechanism which does not involve chelation of a metallic cation. Various other properties of the enzyme are discussed.

Simultaneous Determinations of Liver- and Bone-Type Alkaline Phosphatase by Curve-Fitting of Inhibition Kinetic Data. II. Development and Evaluation of a Fluorescence-Based Method

1992 ◽  
Vol 38 (2) ◽  
pp. 247-255 ◽  
Author(s):  
Carol P Fitzpatrick ◽  
Harry L Pardue
Keyword(s):  
Alkaline Phosphatase ◽  
Kinetic Data ◽  
Kinetic Method ◽  
Linear Least Squares ◽  
First Order ◽  
Least Squares Fit ◽  
Inhibition Kinetic ◽  
Substrate Concentrations ◽  
Measurement Period

Abstract We describe a kinetic method for the determination of alkaline phosphatase (ALP) isoenzymes, based on fluorescence detection of 4-methylumbelliferone. Several different buffer-inhibitor combinations and substrate concentrations were evaluated. Best results were obtained for inhibition with 2.9 mol/L urea in amino-2-methyl-1-propanol buffer. With this combination, normal concentrations of bone- and liver-type ALP could be determined from kinetic data during an 8-min measurement period. We computed initial velocities from parameters for first-order fits during 1.2 half-lives of the response for liver-type ALP. A linear least-squares fit of initial velocities (y) determined in this way vs results obtained with a comparison procedure (x) gave good correlations. We also estimated total signal changes, delta S, from first-order fits during four half-lives. Isoenzyme content correlated well with parameters computed from the first-order fits. Values for standard errors of the estimates represent 5% and 3% of median responses for activities and isoenzyme content, respectively. When compared with an absorbance-based method described previously, this method had threefold shorter measurement times, but imprecisions were 1.6- to 1.8-fold larger.

Interrelationship of the cellular distribution and secretion of regular structure (RS) protein in Bacillus licheniformis nm 105

1992 ◽  
Vol 50 (1) ◽  
pp. 712-713
Author(s):  
Xiaorong Zhu ◽  
Richard McVeigh ◽  
Bijan K. Ghosh
Keyword(s):  
Alkaline Phosphatase ◽  
Bacillus Licheniformis ◽  
Self Assembly ◽  
Gel Electrophoresis ◽  
Culture Supernatant ◽  
Regular Structure ◽  
The Self ◽  
Cellular Distribution ◽  
Structural Protein ◽  
Laboratory Cultures

A mutant of Bacillus licheniformis 749/C, NM 105 exhibits some notable properties, e.g., arrest of alkaline phosphatase secretion and overexpression and hypersecretion of RS protein. Although RS is known to be widely distributed in many microbes, it is rarely found, with a few exceptions, in laboratory cultures of microorganisms. RS protein is a structural protein and has the unusual properties to form aggregate. This characteristic may have been responsible for the self assembly of RS into regular tetragonal structures. Another uncommon characteristic of RS is that enhanced synthesis and secretion which occurs when the cells cease to grow. Assembled RS protein with a tetragonal structure is not seen inside cells at any stage of cell growth including cells in the stationary phase of growth. Gel electrophoresis of the culture supernatant shows a very large amount of RS protein in the stationary culture of the B. licheniformis. It seems, Therefore, that the RS protein is cotranslationally secreted and self assembled on the envelope surface.

In Situ Localization of PPAR Gamma and Uncoupling Protein in Mouse Embryo Sections Using Digoxigenin-Labeled Riboprobes

1996 ◽  
Vol 54 ◽  
pp. 32-33
Author(s):  
C. Jennermann ◽  
S. A. Kliewer ◽  
D. C. Morris
Keyword(s):  
Alkaline Phosphatase ◽  
Room Temperature ◽  
Uncoupling Protein ◽  
Ppar Gamma ◽  
Nuclear Hormone Receptor ◽  
Full Length ◽  
Northern Analysis ◽  
Peroxisome Proliferator

Peroxisome proliferator-activated receptor gamma (PPARg) is a member of the nuclear hormone receptor superfamily and has been shown in vitro to regulate genes involved in lipid metabolism and adipocyte differentiation. By Northern analysis, we and other researchers have shown that expression of this receptor predominates in adipose tissue in adult mice, and appears first in whole-embryo mRNA at 13.5 days postconception. In situ hybridization was used to find out in which developing tissues PPARg is specifically expressed.Digoxigenin-labeled riboprobes were generated using the Genius™ 4 RNA Labeling Kit from Boehringer Mannheim. Full length PPAR gamma, obtained by PCR from mouse liver cDNA, was inserted into pBluescript SK and used as template for the transcription reaction. Probes of average size 200 base pairs were made by partial alkaline hydrolysis of the full length transcripts. The in situ hybridization assays were performed as described previously with some modifications. Frozen sections (10 μm thick) of day 18 mouse embryos were cut, fixed with 4% paraformaldehyde and acetylated with 0.25% acetic anhydride in 1.0M triethanolamine buffer. The sections were incubated for 2 hours at room temperature in pre-hybridization buffer, and were then hybridized with a probe concentration of 200μg per ml at 70° C, overnight in a humidified chamber. Following stringent washes in SSC buffers, the immunological detection steps were performed at room temperature. The alkaline phosphatase labeled, anti-digoxigenin antibody and detection buffers were purchased from Boehringer Mannheim. The sections were treated with a blocking buffer for one hour and incubated with antibody solution at a 1:5000 dilution for 2 hours, both at room temperature. Colored precipitate was formed by exposure to the alkaline phosphatase substrate nitrobluetetrazoliumchloride/ bromo-chloroindlylphosphate.

Electron microscopic study of the membrane glycoproteins in the rat kidney after cisplatin treatment

1989 ◽  
Vol 47 ◽  
pp. 912-913
Author(s):  
S.K. Aggarwal
Keyword(s):  
Alkaline Phosphatase ◽  
Rat Kidney ◽  
Light And Electron Microscopy ◽  
Kidney Tissue ◽  
Membrane Glycoproteins ◽  
Electron Microscopic ◽  
Before And After ◽  
Interstrand Cross Links ◽  
Cross Links

The proposed primary mechanism of action of the anticancer drug cisplatin (Cis-DDP) is through its interaction with DNA, mostly through DNA intrastrand cross-links or DNA interstrand cross-links. DNA repair mechanisms can circumvent this arrest thus permitting replication and transcription to proceed. Various membrane transport enzymes have also been demonstrated to be effected by cisplatin. Glycoprotein alkaline phosphatase was looked at in the proximal tubule cells before and after cisplatin both in vivo and in vitro for its inactivation or its removal from the membrane using light and electron microscopy.Outbred male Swiss Webster (Crl: (WI) BR) rats weighing 150-250g were given ip injections of cisplatin (7mg/kg). Animals were killed on day 3 and day 5. Thick slices (20-50.um) of kidney tissue from treated and untreated animals were fixed in 1% buffered glutaraldehyde and 1% formaldehyde (0.05 M cacodylate buffer, pH 7.3) for 30 min at 4°C. Alkaline phosphatase activity and carbohydrates were demonstrated according to methods described earlier.

The Origin of Elevation of Serum Alkaline Phosphatase in Hepatic Disease

1962 ◽  
Vol 42 (4) ◽  
pp. 431-438 ◽  
Author(s):  
Stanton G. Polin ◽  
Mitchell A. Spellberg ◽  
Lloyd Teitelman ◽  
Makoto Okumura
Keyword(s):  
Alkaline Phosphatase ◽  
Serum Alkaline Phosphatase ◽  
Hepatic Disease

Abstract #208 Paget Disease of Bone with Normal Alkaline Phosphatase

2019 ◽  
Vol 25 ◽  
pp. 67
Author(s):  
Jinetsy Rivera-Ortiz ◽  
Milliette Alvarado-Santiago ◽  
Margarita Ramirez-Vick ◽  
Naomi Collazo-Gutierrez ◽  
Loida Gonzalez-Rodriguez
Keyword(s):  
Alkaline Phosphatase ◽  
Paget Disease Of Bone ◽  
Paget Disease
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