scholarly journals Neutral maltase/glucoamylase from rabbit renal cortex

1989 ◽  
Vol 261 (1) ◽  
pp. 43-47 ◽  
Author(s):  
B Pereira ◽  
S Sivakami
Keyword(s):  
Brush Border ◽  
Brush Border Membrane ◽  
Renal Cortex ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Rabbit Kidney ◽  
Purification Procedure ◽  
Glucoamylase Activity ◽  
Maltase Activity ◽  
Triton X

Maltase activity (EC 3.2.1.20) was solubilized from rabbit kidney brush-border membrane by using 1.0% Triton X-100 and purified 230-fold with an overall recovery of 30%. The purification procedure makes use of heat precipitation, chromatography on DE-52 DEAE-cellulose and gel filtration on Sephacryl S-300. Rabbit kidney brush border exhibited glucoamylase activity with a maltase/glucoamylase ratio of 1.5:1 to 2.0:1. During purification the maltase and glucoamylase activities behaved identically. The Mr of the complex is 590,000, and it appears to be composed of eight identical subunits linked by disulphide bridges.

scholarly journals Alkaline phosphatase from pig kidney. Method of purification and molecular properties

1974 ◽  
Vol 141 (1) ◽  
pp. 273-282 ◽  
Author(s):  
Ernst D. Wachsmuth ◽  
Kunio Hiwada
Keyword(s):  
Alkaline Phosphatase ◽  
Brush Border ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Molecular Properties ◽  
Purification Procedure ◽  
Brush Border Membranes ◽  
Pig Kidney ◽  
Critical Ph

Alkaline phosphatase (EC 3.1.3.1) from pig kidney brush-border membranes was solubilized from membrane precipitates by butan-1-ol at a critical pH of 7.0. The 12000-fold purification procedure included (NH4)2SO4 precipitation, DEAE-and TEAE-cellulose chromatography, Sephadex G-200 gel filtration and neuraminidase digestion followed by DEAE-cellulose chromatography. The purified protein contained 20% (w/w) carbohydrate and had mol.wt. 150000–156000 as estimated by Sephadex filtration and ultracentrifuge analysis. It was a tetrameric glycoprotein consisting of identical subunits, and it had a molecular activity at 25°C of 2600s-1 per tetramer. Its concentration in kidney was estimated to be 8.5–8.8mg/kg.

Some characteristics of kidney Na+ -dependent glucose carrier reconstituted into sonicated liposomes.

1978 ◽  
Vol 234 (1) ◽  
pp. E1
Author(s):  
R K Crane ◽  
P Malathi ◽  
H Preiser ◽  
P Fairclough
Keyword(s):  
Brush Border ◽  
Brush Border Membrane ◽  
Membrane Vesicles ◽  
Rabbit Kidney ◽  
Kidney Cortex ◽  
Triton X ◽  
Glucose Carrier

Studies of the Na+-gradient coupled D-glucose carrier of the brush border membrane of rabbit kidney cortex extracted with Triton X-100 and reconstituted into artificial liposomes show that the reconstituted carrier behaves in those respects so far studied in a manner very similar to the same carrier studied in natural membrane vesicles.

Nicotinic acid transport by brush border membrane vesicles from rabbit kidney

1981 ◽  
Vol 240 (3) ◽  
pp. F185-F191 ◽  
Author(s):  
E. F. Boumendil-Podevin ◽  
R. A. Podevin
Keyword(s):  
Membrane Potential ◽  
Nicotinic Acid ◽  
Brush Border ◽  
Brush Border Membrane ◽  
Isonicotinic Acid ◽  
Membrane Vesicles ◽  
Brush Border Membrane Vesicles ◽  
Rabbit Kidney ◽  
Internal Space ◽  
Acid Uptake

The transport of nicotinic acid was investigated in brush border membrane vesicles isolated from rabbit kidney. The imposition of a Na+ gradient (out to in) induced a transient stimulation of nicotinic acid uptake above its final equilibrium value. This stimulation was specific for Na+. The uptake of nicotinic acid by the brush border membranes represented transport into an internal space and occurred in the absence of significant nicotinic acid degradation. The Na+ gradient-dependent uptake of nicotinic acid was saturable, apparent Km = 0.3 mM. Uptake of nicotinic acid was inhibited by its two isomers: picolinic and isonicotinic acid. In contrast, pyridine derivatives with two carboxyl groups or an amide group in addition to the carboxyl group were without inhibitory effect. Evaluation of changes in membrane potential using the lipophilic cation triphenylmethylphosphonium demonstrated that conditions that transiently generated either an interior-positive or an interior-negative membrane potential failed to affect the Na+-dependent transport of nicotinic acid. These findings provide evidence of the existence on the luminal membrane of a Na+ gradient-dependent and electroneutral transport system for nicotinic acid.

scholarly journals Characterization of a Novel Plasma Membrane Protein, Expressed in the Midgut Epithelia of Bombyx mori, That Binds to Cry1A Toxins

2004 ◽  
Vol 70 (8) ◽  
pp. 4604-4612 ◽  
Author(s):  
Delwar M. Hossain ◽  
Yasuyuki Shitomi ◽  
Kenta Moriyama ◽  
Masahiro Higuchi ◽  
Tohru Hayakawa ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Brush Border ◽  
Membrane Vesicle ◽  
Gel Filtration ◽  
Resistance Mechanism ◽  
Binding Assays ◽  
Toxin Resistance ◽  
Cry Toxin ◽  
Triton X

ABSTRACT We describe the properties of a novel 252-kDa protein (P252) isolated from brush border membranes of Bombyx mori. P252 was found in a Triton X-100-soluble brush border membrane vesicle fraction, suggesting that it may be a component of the midgut epithelial cell membrane. P252 was purified to homogeneity, and the amino acid sequence of two internal peptides was determined, but neither of the peptides matched protein sequences in the available databases. The apparent molecular mass of the purified protein was estimated by denaturing gel electrophoresis to be 252 kDa, and it migrated as a single band on native gels. However, gel filtration chromatography indicated an apparent mass of 985 kDa, suggesting that P252 may exist as a homo-oligomer. The associations of P252 with Cry1Aa, Cry1Ab, and Cry1Ac were specific, and Kd constants were determined to be 28.9, 178.5, and 20.0 nM, respectively. A heterologous competition assay was also done. P252 did not exhibit Leu-pNA hydrolysis activity, and binding to the Cry1A toxins was not inhibited by GalNAc. Binding assays of P252 with various lectins indicated the presence of three antennal N-linked high-mannose-type as well as O-linked mucin-type sugar side chains. While the function of P252 is not yet clear, we propose that it may function with Cry1A toxins during the insecticidal response and/or Cry toxin resistance mechanism.

Effect of thyroxine administration on phosphate transport across renal cortical brush border membrane

1984 ◽  
Vol 246 (2) ◽  
pp. F133-F139 ◽  
Author(s):  
R. E. Espinosa ◽  
M. J. Keller ◽  
A. N. Yusufi ◽  
T. P. Dousa
Keyword(s):  
Brush Border ◽  
Brush Border Membrane ◽  
Renal Cortex ◽  
Membrane Vesicles ◽  
Phosphate Transport ◽  
Treated Group ◽  
Renal Tubular Reabsorption ◽  
Renal Tubular ◽  
And Control ◽  
Cortical Slices

Previous studies indicate that in hyperthyroid states the renal tubular reabsorption of phosphate is enhanced. To determine the cellular basis of this phenomenon, we investigated the effect of L-thyroxine (T4) administration on Pi transport across brush border membrane vesicles (BBMV) from rat renal cortex. Rats were thyroparathyroidectomized, fed a diet containing 1.2% phosphate, and treated intraperitoneally for 6 days with 200 micrograms T4 X 100 g body wt-1 X day-1. At the end of the treatment period, the rats had a significantly (+ delta 25%) elevated plasma Pi and a slightly decreased plasma Ca compared with controls. The renal clearance of Pi was not different between the two groups. Na+ gradient-dependent uptake of 32Pi by BBMV from renal cortex was significantly enhanced in T4-treated rats. BBMV uptake of 32Pi in the absence of Na+ -gradient as well as Na+ gradient-dependent uptake of D-[3H]glucose and L-[3H]proline did not differ between BBMV from T4-treated and control rats. Kinetic analysis showed that the Na+ gradient-dependent 32Pi transport system in BBMV from T4-treated rats had a significantly increased Vmax compared with controls (5.2 +/- 0.4 vs. 4.1 +/- 0.4 nmol Pi X 30 s-1 X mg protein-1) and also a slightly higher affinity for Pi (apparent Km in controls, 95 +/- 9; in T4-treated, 78 +/- 8 microM). Gluconeogenesis in cortical slices was not significantly different between T4-treated rats and controls. Specific activities of alkaline phosphatase and gamma-glutamyltransferase were significantly lower in BBMV from the T4-treated group compared with controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Na+-dependent neutral amino acid transporter ATB0 is a rabbit epithelial cell brush-border protein

2001 ◽  
Vol 281 (3) ◽  
pp. C963-C971 ◽  
Author(s):  
Nelly E. Avissar ◽  
Charlotte K. Ryan ◽  
Vadivel Ganapathy ◽  
Harry C. Sax
Keyword(s):  
Amino Acid ◽  
Hela Cells ◽  
Brush Border ◽  
Brush Border Membrane ◽  
Polyclonal Antibodies ◽  
Neutral Amino Acid ◽  
Rabbit Kidney ◽  
Western Blots ◽  
Amino Acid Transport System ◽  
Rabbit Tissues

System B0 activity accounts for the majority of intestinal and kidney luminal neutral amino acid absorption. An amino acid transport system, called ATB0 (also known as ASCT2), with functional characteristics similar to those of system B0, has been recently cloned. We generated polyclonal antibodies to human and rabbit ATB0 COOH-terminal peptides and used Western blot analysis to detect ATB0 protein in rabbit tissues, rabbit ileal brush-border membrane vesicles (BBMV), and HeLa cells transfected with plasmids containing ATB0 cDNAs. Immunohistochemistry was used to localize ATB0 in rabbit kidney and intestine. In Western blots of rabbit tissues, ATB0 was a broad smear of 78- to 85-kDa proteins. In transfected HeLa cells, ATB0 appeared as a smear consisting of 57- to 65-kDa proteins. The highest expression was found in the kidney. ATB0 was enriched in rabbit ileal BBMV and in HeLa cells transfected with ATB0 cDNAs. In the kidney and in the intestine, ATB0 was confined to the brush-border membrane (BBM) of the proximal tubular cell and of the enterocyte, respectively. Tissue and intracellular distribution of ATB0 protein parallels that of system B0 activity. ATB0protein could be the transporter responsible for system B0in the BBM of epithelial cells.

scholarly journals ISOLATION AND BIOCHEMICAL CHARACTERIZATION OF BRUSH BORDERS FROM RABBIT KIDNEY

1970 ◽  
Vol 47 (3) ◽  
pp. 637-645 ◽  
Author(s):  
Sosamma J. Berger ◽  
Bertram Sacktor
Keyword(s):  
Brush Border ◽  
Renal Cortex ◽  
Biochemical Characterization ◽  
Rabbit Kidney ◽  
Renal Tubules ◽  
Marker Enzymes ◽  
Alkaline Phosphatases ◽  
Brush Borders ◽  
Specific Activities

A technique for the isolation of intact brush borders from rabbit renal cortex was evaluated. The procedure was monitored by phase and electron microscopy and marker enzymes, i.e. ATP:NMN adenylyl transferase, nuclear; cytochrome oxidase, mitochondrial; ß-glucuronidase, lysosomal; and glucose-6-Pase, microsomal; and indicated an essentially pure preparation of brush borders. The disaccharidase, trehalase, previously reported in renal tubules, was localized uniquely in brush borders. Maltase was also found; the specific activities of the two enzymes in the brush borders were increased 10- to 20-fold. Other disaccharidases, such as sucrase, isomaltase, lactase, and cellobiase, were absent. It is suggested that trehalase and maltase are appropriate candidates for marker enzymes of the renal brush border. Isolated brush borders possessed a ouabain-sensitive (Na+ + K+) ATPase, an oligomycin-insensitive Mg++ ATPase, and a Ca++-activated ATPase. Alkaline phosphatases, dephosphorylating ß-glycero-P, and trehalose-6-P were also present. The specific activities of these enzymes were increased three-to-five fold in the brush-border preparations; however, activities were found in other subcellular fractions of the renal cortex. Hexokinase, although evident in the isolated brush border, was found prominently associated with other membranous fractions. Phosphoglucomutase and UDPG pyrophosphorylase were localized in the soluble fraction of the renal cortex.

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