Electrophoretic variants of alanine aminopeptidase, alkaline phosphatase, and gamma-glutamyltransferase in urine.

1984 ◽  
Vol 30 (6) ◽  
pp. 856-859 ◽  
Author(s):  
K Jung ◽  
U W Wischke
Keyword(s):  
Alkaline Phosphatase ◽  
Electrophoretic Mobility ◽  
Brush Border ◽  
Neuraminidase Treatment ◽  
Electrophoretic Variants ◽  
Brush Border Enzymes ◽  
Gamma Glutamyltransferase ◽  
Electrophoretic Mobilities ◽  
Alanine Aminopeptidase ◽  
Triton X

Abstract The three brush-border enzymes--alanine aminopeptidase (EC 3.4.11.2), alkaline phosphatase (EC 3.1.3.1), and gamma-glutamyltransferase (EC 2.3.2.2)--are present in the urine of healthy persons in two variants, a particulate form and a soluble one. They can be separated by electrophoresis in agarose gel and by ultracentrifugation. The particulate forms exhibit similar electrophoretic mobility, but the soluble forms of these brush-border enzymes differ in their electrophoretic mobilities. The enzyme components of the particulate activity can be mobilized by Triton X-100 and trypsin. The electrophoretic mobility of the soluble forms of alanine aminopeptidase and gamma-glutamyltransferase is slowed by neuraminidase treatment. Both forms of gamma-glutamyltransferase are influenced in their electrophoretic mobility by treatment with n-butanol/diisopropyl ether, showing their lipid dependence. These findings enhance our knowledge of the biochemical nature of brush-border enzymes in urine.

Different diuresis-dependent excretions of urinary enzymes: N-acetyl-beta-D-glucosaminidase, alanine aminopeptidase, alkaline phosphatase, and gamma-glutamyltransferase.

1986 ◽  
Vol 32 (3) ◽  
pp. 529-532 ◽  
Author(s):  
K Jung ◽  
G Schulze ◽  
C Reinholdt
Keyword(s):  
Alkaline Phosphatase ◽  
Brush Border ◽  
Excretion Rate ◽  
Urine Flow ◽  
High Intake ◽  
Brush Border Enzymes ◽  
Urinary Creatinine ◽  
Gamma Glutamyltransferase ◽  
Alanine Aminopeptidase ◽  
Brush Border Enzyme

Abstract We studied how much of the lysosomal enzyme N-acetyl-beta-D-glucosaminidase (EC 3.2.1.30) and of the brush-border enzymes alanine aminopeptidase (EC 3.4.11.2), alkaline phosphatase (EC 3.1.3.1), and gamma-glutamyltransferase (EC 2.3.2.2) was excreted in urine over 8 h after a high intake of fluid (22 mL per kilogram of body weight). The hourly excretion of all four enzymes increased with the increasing urine flow rate. The excretion rate of the brush-border enzymes was more markedly influenced than that of N-acetyl-beta-D-glucosaminidase. By relating the enzyme excretion to urinary creatinine we could reduce the variability of brush-border enzyme output and could completely compensate for the effect of diuresis on the excretion of N-acetyl-beta-D-glucosaminidase.

Diagnostic significance of some urinary enzymes for detecting acute rejection crises in renal-transplant recipients: alanine aminopeptidase, alkaline phosphatase, gamma-glutamyltransferase, N-acetyl-beta-D-glucosaminidase, and lysozyme.

1986 ◽  
Vol 32 (10) ◽  
pp. 1807-1811 ◽  
Author(s):  
K Jung ◽  
J Diego ◽  
V Strobelt ◽  
D Scholz ◽  
G Schreiber
Keyword(s):  
Alkaline Phosphatase ◽  
Renal Transplant ◽  
Acute Rejection ◽  
Renal Transplant Recipients ◽  
Transplant Recipients ◽  
Urinary Enzymes ◽  
Gamma Glutamyltransferase ◽  
Alanine Aminopeptidase ◽  
Group A ◽  
Group B

Abstract We compared the diagnostic validity of five urinary enzymes--alanine aminopeptidase (EC 3.4.11.2), alkaline phosphatase (EC 3.1.3.1), gamma-glutamyltransferase (EC 2.3.2.2), N-acetyl-beta-D-glucosaminidase (EC 3.2.1.30), and lysozyme (EC 3.2.1.17)--as indicators of acute rejection crises in renal-transplant recipients. In 82 patients (group A), the excretion of each of these five enzymes was measured daily from transplantation until discharge from hospital. In another 69 patients (group B), enzyme determinations were made when the patient came for regular checkups (about every four to eight weeks). We used an "activity ratio" (the activity measured at a particular time compared with the activity on the preceding determination) value of 1.5 as the decision point. In group A, use of this discrimination point for alanine aminopeptidase, gamma-glutamyltransferase, and N-acetyl-beta-D-glucosaminidase yielded a specificity and sensitivity of about 90%. In group B, only alanine aminopeptidase had a greater diagnostic sensitivity than creatinine alone. Evidently, measurement of alanine aminopeptidase can be a helpful indicator of acute rejection crises, when interpreted in combination with other available relevant clinical, biochemical, and immunological data.

Improved agarose electrophoretic method for separating alkaline phosphatase isoenzymes in serum.

1988 ◽  
Vol 34 (9) ◽  
pp. 1857-1862 ◽  
Author(s):  
V O Van Hoof ◽  
L G Lepoutre ◽  
M F Hoylaerts ◽  
R Chevigné ◽  
M E De Broe
Keyword(s):  
Alkaline Phosphatase ◽  
Monoclonal Antibodies ◽  
Polyclonal Antibody ◽  
High Molecular Mass ◽  
High Reproducibility ◽  
Neuraminidase Treatment ◽  
Electrophoretic Method ◽  
Electrophoretic Mobilities ◽  
Before And After ◽  
Alkaline Phosphatase Isoenzymes

Abstract A modified agarose electrophoretic system for the separation of alkaline phosphatase (ALP, EC 3.1.3.1) isoenzymes is described. Bone, liver, high-molecular-mass, and intestinal ALP are separated with high reproducibility. The sensitivity of the agarose system is superior to cellulose acetate in detecting high-Mr ALP. Correlation is good between bone ALP fractions scanned before and after treatment with neuraminidase. Immunoglobulin-bound ALPs, the ALP-lipoprotein-X complex, and the additional ALP fraction observed in transient hyperphosphatasemia in children are detected by their peculiar electrophoretic mobility in the proposed system. Approximately 25% of the samples contained an additional fraction of intestinal-type ALP, as evidenced by neuraminidase treatment and use of polyclonal and monoclonal antibodies. Because the electrophoretic mobilities of this "intestinal variant" and of some immunoglobulin-bound ALP fractions are identical to those of bone and intestinal ALP, respectively, treatment of the samples with a polyclonal antibody that reacts with intestinal ALP is advised.

scholarly journals Characterization of neutral endopeptidase 24.11 in dog glomeruli

1993 ◽  
Vol 291 (3) ◽  
pp. 773-779 ◽  
Author(s):  
C Landry ◽  
P Santagata ◽  
W Bawab ◽  
M C Fournié-Zaluski ◽  
B P Roques ◽  
...  
Keyword(s):  
Cell Surface ◽  
Brush Border ◽  
Neutral Endopeptidase ◽  
Glycosylation Pattern ◽  
Electrophoretic Mobilities ◽  
Mammalian Tissues ◽  
Dog Kidney ◽  
A Cell ◽  
Membrane Bound ◽  
Triton X

Neutral endopeptidase (NEP; also known as neprilysin and enkephalinase; EC 3.4.24.11) is a cell-surface metallopeptidase that is present in many mammalian tissues. It is particularly abundant on the brush-border membranes of the kidney proximal tubule. In this paper, the presence of NEP in purified glomeruli from dog kidney was assessed by measuring phosphoramidon- and thiorphan-sensitive [D-Ala2,Leu5]enkephalin-degrading activity. Using this assay, the Km and kcat. of the glomerular enzyme were found to be identical to those of the tubular enzyme. By Western blotting the apparent M(r) of the glomerular enzyme was found to be 104,000, compared with 94,000 for the tubular enzyme. This might be due to a different glycosylation pattern, since endoglycosidase F treatment of NEP obtained from both tissues yielded deglycosylated enzymes with similar electrophoretic mobilities. The glomerular enzyme also appears to be membrane-bound, since it was retained in the detergent-rich phase after phase separation with Triton X-114. Autoradiography experiments performed with RB104, a new highly selective and potent NEP inhibitor, showed that NEP was expressed in both glomeruli and proximal tubules. The presence in glomeruli of NEP and some other brush-border peptidases (dipeptidyl-dipeptidase IV, aminopeptidase N and angiotensin I-converting enzyme) suggests that cell-surface peptidases might play an important role as regulators of plasma-derived peptides in this part of the nephron.

scholarly journals Plasma lipopolysaccharide level and enterocyte brush border enzymes in gnotobiotic piglets infected with Salmonella typhimurium

2012 ◽  
Vol 47 (No. 10 - 11) ◽  
pp. 289-294
Author(s):  
I. Trebichavský ◽  
H. Kozáková ◽  
IŠplíchal
Keyword(s):  
Alkaline Phosphatase ◽  
Brush Border ◽  
Virulent Strain ◽  
Systemic Infection ◽  
Gamma Glutamyl Transpeptidase ◽  
Brush Border Enzymes ◽  
Dipeptidylpeptidase Iv ◽  
Glutamyl Transpeptidase ◽  
Rough Mutant ◽  
Gnotobiotic Piglets

Gnotobiotic piglets were orally infected either with the virulent LT2 strain or the non-pathogenic SF1591 rough mutant of Salmonella enterica serotype Typhimurium. They were sacrificed 6 or 24 h after the infection. All piglets infected for 24 h developed systemic infection with an increase of plasma lipopolysaccharide. Infection with the virulent strain caused a significant decrease (P < 0.001) of gamma-glutamyl transpeptidase (GGT) activity in the enterocyte brush border of both the jejunum and ileum, infection with the rough mutant caused a decrease of GGT activity in the ileum only. The activities of other brush border enzymes (lactase, sucrase, glucoamylase, alkaline phosphatase and dipeptidylpeptidase IV) did not change significantly after infection.

High-molecular-mass alkaline phosphatase in serum and bile: nature and relationship with lipoprotein-X.

1981 ◽  
Vol 27 (6) ◽  
pp. 867-874 ◽  
Author(s):  
P M Crofton ◽  
A F Smith
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Molecular Mass ◽  
Electrophoretic Mobility ◽  
Leucine Aminopeptidase ◽  
Current Knowledge ◽  
Normal Serum ◽  
High Molecular Mass ◽  
Variable Effect ◽  
Gamma Glutamyltransferase

Abstract Using immunoelectrophoresis and other techniques, we have demonstrated an association between lipoprotein-X and (a) alkaline phosphatase and (b) other enzymes originating from the hepatocyte membrane, namely gamma-glutamyltransferase and leucine aminopeptidase. The high-molecular-mass forms of these enzymes, in both serum and bile, were precipitated by lipoprotein-X antiserum but not by antisera to other plasma proteins. The activity of high-molecular-mass alkaline phosphatase in serum was positively correlated with lipoprotein-X and with lipoprotein-X-associated alkaline phosphatase, both assessed semi-quantitatively. On the other hand, many sera possessed high activities of high-molecular-mass alkaline phosphatase but no detectable lipoprotein-X. Incubation of serum with conjugated bile salts and with synthetic detergents, at concentrations which did not dissociate the high-molecular-mass enzymes, caused parallel alterations in the electrophoretic mobility of serum lipoprotein-X and its associated enzyme activity. Incubation of normal dialyzed hepatic bile with normal, lipoprotein-X-negative serum produced an alteration in electrophoretic mobility of biliary lipoprotein and its associated enzyme activity from anodal to cathodal in agar gel. Digestion with papain had a variable effect on the different enzymes in the complex, without affecting the lipoprotein moiety. Leucine aminopeptidase was removed most readily from the complex to give the low-molecular-mass form present in normal serum; gamma-glutamyltransferase dissociated somewhat less readily, and alkaline phosphatase was completely resistant to dissociation from the complex. These results are discussed in the light of current knowledge, and a hypothesis is proposed for the nature of the high-molecular-mass enzymes in serum and bile.

Surface Modification and Electrophoresis of Normal and Transformed BHK21 Cells

1974 ◽  
Vol 14 (1) ◽  
pp. 203-214
Author(s):  
A. L. LATNER ◽  
G. A. TURNER
Keyword(s):  
Electrophoretic Mobility ◽  
Cell Types ◽  
Transformed Cells ◽  
Normal Cells ◽  
Neuraminidase Treatment ◽  
Total Histone ◽  
Electrophoretic Mobilities ◽  
Formaldehyde Treatment ◽  
The Mean ◽  
Number Of Cells

The mean electrophoretic mobilities at pH 7.5 of virus-transformed (Py6) and normal BHK21 cells were very similar, whether they were harvested mechanically or by the use of trypsin. After formaldehyde treatment, there was significantly increased mobility in both cell types; the transformed cells showed significantly the greater change. After neuraminidase treatment, the mean electrophoretic mobility was decreased to the same extent in both types of cell. Treatment with neuraminidase and formaldehyde had no effect on the mean electrophoretic mobility of the normal cells but slowed the transformed variety. The mobility in histone solution had no relationship to histone concentration but was statistically correlated with the amount of histone per cell, calculated from total histone present divided by the total number of cells; a linear relationship being obtained with the normal cells but an initial plateau was demonstrated with the transformed cells. The normal cell line showed a similar plateau after neuraminidase treatment. The possible significance of these results is discussed.

Effects of genetic strain and holding facility on the characteristics of alkaline phosphatase and brush border enzymes in silver perch (Bidyanus bidyanus)

2007 ◽  
Vol 38 (4) ◽  
pp. 361-372 ◽  
Author(s):  
Yaniv Hakim ◽  
Stuart J Rowland ◽  
Jeff A Guy ◽  
Charles Mifsud ◽  
Zehava Uni ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Brush Border ◽  
Brush Border Enzymes ◽  
Genetic Strain

Biochemical and ultrastructural investigation of the effect of Stelazine (trifluoperazine) on Hymenolepis diminuta (Cestoda)

Parasitology ◽  
1987 ◽  
Vol 94 (1) ◽  
pp. 135-149 ◽  
Author(s):  
Jayne B. Hipkiss ◽  
A. Skinner ◽  
C. J. Branford White
Keyword(s):  
Alkaline Phosphatase ◽  
Connective Tissue ◽  
Brush Border ◽  
Incubation Medium ◽  
Hymenolepis Diminuta ◽  
Minimal Effect ◽  
Cellular Organization ◽  
Brush Border Enzymes ◽  
Ultrastructural Appearance

SUMMARYThe effects of the phenothiazine, Stelazine, on Hymenolepis diminuta were investigated. The cestode was incubated for 10 min at 37 °C with 1 mM trifluoperazine, in the presence and absence of Ca2+. Assay of brush border enzymes showed that drug treatment lowered the activities of alkaline phosphatase, Ca2+-ATP'ase, 5′-nucleotidase and type 1 phosphodiesterase. This occurred in parallel with a significant reduction in tegumental protein. Under these conditions gross changes in ultrastructural appearance and cellular organization were observed. There was a lack of ordered microtriches and the distal cytoplasm was absent. Glycogen granules were scattered throughout the cytoplasm within the subtegumental layer. The connective tissue also appeared to be in some disarray. The effects of Stelazine appeared to be dependent on time and were significantly increased when Ca2+ was included in the incubation medium. Incubation with the less hydrophobic phenothiazine trifluoperazine sulphoxide had minimal effect on the integrity of the cestode. The results reported here support the premise that certain phenothiazines may be considered as potential cestocidal agents.

Sign in / Sign up

Export Citation Format

Share Document