Aluminum-Binding Protein in Dialysis Dementia. I. Characterization in Plasma by Gel Chromatography and Electrophoresis

Nephron ◽  
1989 ◽  
Vol 52 (4) ◽  
pp. 323-328 ◽  
Author(s):  
Farhad Khalil-Manesh ◽  
Cathy Agness ◽  
Harvey C. Gonick
Keyword(s):  
Binding Protein ◽  
Gel Chromatography ◽  
Dialysis Dementia

scholarly journals Affinity purification of cellular retinoic acid-binding protein on 14-carboxy-13-cis-retinamide-sepharose 4B

1989 ◽  
Vol 262 (3) ◽  
pp. 917-922 ◽  
Author(s):  
R K Singh ◽  
B P Sani ◽  
M I Dawson ◽  
Y F Shealy
Keyword(s):  
Retinoic Acid ◽  
Carboxy Group ◽  
Binding Proteins ◽  
Binding Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Biologically Active ◽  
Amide Bond ◽  
Gel Chromatography ◽  
Fatty Acid Binding ◽  
Sepharose 4B

A biologically active bifunctional retinoid, ethyl 14-carboxyretinoate, has been synthesized and shown to bind cellular retinoic acid (RA)-binding protein (CRABP) via its free carboxy group. We describe herein the synthesis of 14-carboxy-13-cis-retinamide-Sepharose 4B, which is an affinity matrix bearing an all-trans-RA moiety, and thus was used to purify and characterize CRABP from chick-embryo skin. An amide bond was first formed between the free carboxy group of the retinoid and a primary amino group of aminohexyl-Sepharose 4B, by reaction with carbodi-imide, and the ester group of the resin-bound retinoid was then hydrolysed in an alkaline medium. Polyacrylamide-gel electrophoresis and f.p.l.c. Superose column-chromatographic analysis demonstrated that the affinity-purified CRABP (Mr 15,000) was close to electrophoretic homogeneity (greater than 90%) and specifically interacts with RA. By using affinity gel chromatography, conversion of holo-CRABP into apo-CRABP by treatment with p-hydroxymercuribenzoate and a possible involvement of a thiol group in RA binding to CRABP were established. This affinity procedure provides several advantages: (i) 14-carboxy-13-cis-retinamide-Sepharose exhibited high efficiency and selectivity for RA-binding protein (i.e. retinol- or fatty-acid-binding proteins did not bind); (ii) the presence of the amide linkage between the ligand and the matrix makes this affinity resin relatively stable to cytosolic enzymes; and (iii) other RA-binding proteins, e.g. nuclear receptor(s), may be purified.

High-affinity folate binding in human prostate

1993 ◽  
Vol 13 (2) ◽  
pp. 99-105 ◽  
Author(s):  
Jan Holm ◽  
Steen Ingemann Hansen ◽  
Mimi Høier-Madsen
Keyword(s):  
Human Milk ◽  
Binding Protein ◽  
Cross Reactivity ◽  
Human Prostate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Gel Chromatography ◽  
Folate Binding Protein ◽  
High Affinity ◽  
Sds Page ◽  
Triton X

Binding of 3H-folate in Triton X-100 solubilized human prostate homogenate was of a high-affinity type and displayed apparent positive cooperativity typical of specific folate binding. Radioligand dissociation was slow at pH 7.4, but rapid at pH 3.5. Gel chromatography reveled two major folate binding proteins (Mr≈100 and 25kDa), but only one single band (Mr ≈ 65–70 kDa) was detectable on SDS-PAGE and immunoblotting with rabbit-anti human milk folate binding protein. Concentration of folate binding protein in prostate homogenate expressed as maximum 3H-folate binding was 1.10 nmol/g protein, and the cross-reactivity with rabbit-anti human milk folate binding protein serum was 15% as determined by an enzyme-linked immunosorbent assay (median values; n = 6).

Growth hormone binding protein in patients with renal failure

1992 ◽  
Vol 127 (6) ◽  
pp. 485-488 ◽  
Author(s):  
Hiralal G Maheshwari ◽  
Ian Rifkin ◽  
Joan Butler ◽  
Michael Norman
Keyword(s):  
Growth Hormone ◽  
Renal Failure ◽  
Renal Disease ◽  
Binding Protein ◽  
Scatchard Analysis ◽  
Gel Chromatography ◽  
Hormone Binding ◽  
Binding Studies ◽  
Growth Hormone Binding Protein ◽  
Hormone Binding Protein

To investigate changes in the growth hormone binding protein (GH-BP) in renal disease, gel chromatography was used to separate free and bound hormone after incubation with 125I-GH, the results being expressed as a percentage of radioactive GH eluting in a high molecular weight (70–80 kD) peak. In 26 normal individuals, binding was 39.3±8.0%, while in 11 patients with renal disease who were off dialysis binding was reduced to 16.8±5.6%. Similarly, in 9 patients undergoing hemodialysis binding was reduced to 24.6±6.8%, in 8 patients undergoing chronic ambulatory peritoneal dialysis binding was reduced to 25.7±7.6%, and in 9 patients within three months of a renal transplant binding was reduced to 25.1±8.6%. Scatchard analysis showed that these changes were not a result of decreased affinity of GH-BP for GH, and receptor binding studies showed that uremic serum was not inhibiting binding. The decreased concentration of GH-BP may indicate decreased expression of the GH receptor in target tissues, and hence diminished responsiveness to GH in renal failure.

Aluminum-Binding Protein in Dialysis Dementia. II. Characterization in Plasma by Ultrafiltration

Nephron ◽  
1989 ◽  
Vol 52 (4) ◽  
pp. 329-333 ◽  
Author(s):  
Farhad Khalil-Manesh ◽  
Cathy Agness ◽  
Harvey C. Gonick
Keyword(s):  
Binding Protein ◽  
Dialysis Dementia

Release of insulin-like growth factor I by the sheep placenta in vitro

1991 ◽  
Vol 3 (4) ◽  
pp. 379 ◽  
Author(s):  
J Falconer ◽  
JJ Davies ◽  
HP Zhang ◽  
R Smith
Keyword(s):  
Molecular Weight ◽  
Growth Factor ◽  
Binding Protein ◽  
Basal Plate ◽  
Gel Chromatography ◽  
Insulin Like Growth Factor ◽  
Factor I ◽  
Igf I ◽  
Growth Factor I ◽  
Villous Tissue

This study has utilized a tissue perifusion system to examine the release of insulin-like growth factor I (IGF I) from different regions of the sheep placenta at two stages of gestation. Placentae were obtained from ewes at either 116 +/- 7 (mean +/- s.e.) or 144 +/- 1.5 days of gestation and separated into the maternal basal plate or chorionic villous tissue. At both ages, the maternal basal plate tissue released approximately three times more IGF I than the chorionic villous tissue. No difference was found between the rate of release of IGF I from the maternal basal plate at 120 and 140 days of gestation, whereas the chorionic villous tissue released less IGF I later in gestation. Maternal basal plate tissue was less responsive to a depolarizing dose of KCl than was chorionic villous tissue at either age. After acid gel chromatography, perfusate from the basal plate had three peaks of IGF immunoreactivity (corresponding to binding protein, IGF I and a form with an intermediate molecular weight). In contrast, the chorionic villous tissue released only a form with a high molecular weight, corresponding to binding protein. These results demonstrate that the sheep placenta produced IGF I, that secretion varies between different placental zones which contain different cell types and that there are maturational changes in placental IGF I secretion. The IGFs may be involved in placental growth.

High-affinity folate binding in human mammary gland

1993 ◽  
Vol 13 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Jan Holm ◽  
Steen Ingemann Hansen ◽  
Mimi Høier-Madsen
Keyword(s):  
Mammary Gland ◽  
Binding Protein ◽  
Cross Reactivity ◽  
Binding Activity ◽  
Gel Chromatography ◽  
Folate Binding Protein ◽  
High Affinity ◽  
Mammary Gland Tissue ◽  
Human Mammary Gland ◽  
Triton X

High-affinity 3H-folate binding in Triton X-100 solubilized human mammary gland tissue displayed characteristics, e.g. apparent positive cooperativity and increasing affinity with decreasing concentration of folate binding protein, shown to be typical of specific folate binding. Radioligand dissociation was slow at pH 7.4. A major fraction of the bound radioligand dissociated rapidly at pH 3.5, while a residual binding of 20% persisted even after prolonged dialysis at pH 3.5. Gel chromatography revealed two major folate binding proteins (Mr ≈ 100 kDa and 25 kDa). However, only one single band was detectable on SDS-PAGE immunoblotting. The highest folate binding activity per g protein was associated with the upper triglyceride-containing layer of the 1000 g supernatant of the homogenate. The folate binding protein extracted from this layer had a low cross-reactivity (<5%) with rabbit antibodies against 25 kDa human milk folate binding protein. The folate binding protein in the 1000 g pellet and the aqueous phase of the 1000 g supernatant was present at a low concentration and had a cross-reactivity of 100%.

scholarly journals Purification and characterization of a cholesterol-binding protein from human pancreas

1982 ◽  
Vol 207 (3) ◽  
pp. 573-582 ◽  
Author(s):  
A Sziegoleit
Keyword(s):  
Serum Proteins ◽  
Binding Capacity ◽  
Binding Protein ◽  
Protein Composition ◽  
Sodium Deoxycholate ◽  
Gel Chromatography ◽  
Human Pancreas ◽  
Lipid Complex ◽  
Intestinal Fluids ◽  
Cholesterol Binding

The protein composition of human intestinal lavage fluids was analysed by electroimmunoassay. In addition to secretory immunoglobulin A and other components that were antigenically related to serum proteins, a number of gut-specific proteins were detected. One of these was found to exhibit the capacity of binding sodium deoxycholate and cholesterol. After isolation of this cholesterol-binding protein from intestinal fluids, immunohistochemical studies utilizing a specific antiserum indicated the pancreas to be the organ of its synthesis. The protein was subsequently purified from necrobiotic pancreas tissues and was found to be composed of a single polypeptide chain with a mol. wt. of 28 000 and an isoelectric point of pH4.9. The deoxycholate binding capacity determined by gel chromatography in the presence of [3H]deoxycholate was calculated to be approx. 24 mol of deoxycholate/mol of protein. In the intestinal fluids the protein appeared to be present in firm association with cholesterol, phospholipids, triacylglycerols and bile salts as a macromolecular protein-lipid complex. The possibility is raised that the pancreas-derived, cholesterol-binding protein may fulfil a function as an intestinal ‘lipoprotein’.

Cross-reaction of serum somatomedin-binding protein in a radioimmunoassay developed for somatomedin-binding protein isolated from human amniotic fluid

1984 ◽  
Vol 107 (4) ◽  
pp. 563-570 ◽  
Author(s):  
Guilherme Póvoa ◽  
Arved Roovete ◽  
Kerstin Hall
Keyword(s):  
Amniotic Fluid ◽  
Molecular Form ◽  
Binding Protein ◽  
Molecular Size ◽  
Diabetic Patients ◽  
Gel Chromatography ◽  
Human Amniotic Fluid ◽  
Response Curves ◽  
Protein Gel ◽  
The Mean

Abstract. A radioimmunoassay was developed for somatomedin-binding protein isolated from human amniotic fluid. The mean level in amniotic samples from 20–22 week of gestation was high, 51 μg/ml. Human serum and serum fraction gave dose-response curves superimposable on that for the pure amniotic binding protein. Gel chromatography of serum at neutral pH disclosed immunoreactive binding protein only in fractions with a molecular size of 35000 corresponding to elution volume for the low molecular form somatomedin-binding protein. The mean levels (mean and range) of immunoreactive somatomedin-binding protein in cord blood (191 ng/ml, 55–1698 ng/ml) and in diabetic patients with uraemia (97 and 51–174 ng/ml) were 5- to 10-fold elevated above those of healthy adults (23 and 18–36 ng/ml). In acromegaly the levels decreased with increasing GH production (r = −0.77; P < 0.005). In adult patients with GH deficiency the levels were 2-fold elevated in comparison with healthy subjects. Apart from patients with uraemia a negative correlation was found in adults between the levels of immunoreactive binding protein and immunoreactive IGF-I which reflect the GH production (r = −0.81, P < 0.001).

Immunoreactive insulin-like growth factor II in urine

1990 ◽  
Vol 123 (5) ◽  
pp. 499-503 ◽  
Author(s):  
Walter Zumkeller ◽  
Kerstin Hall
Keyword(s):  
Growth Factor ◽  
Binding Protein ◽  
Immunoreactive Insulin ◽  
Gel Chromatography ◽  
Insulin Like Growth Factor ◽  
Factor Ii ◽  
Igf I ◽  
Chromatographic Profile ◽  
Igf Binding ◽  
Igf Binding Protein

Abstract. Insulin-like growth factor II and insulin-like growth factor binding protein-1 were identified and quantified in the urine of 23 healthy subjects between 17 and 76 years of age. IGF-II was measured after separation by gel chromatography at low pH and compared with IGF-I levels in the same samples, whereas IGF binding protein-1 was measured in dialysed urine. Urinary IGF-II was found at much higher concentrations than IGF-I (mean ±sem: 717±69 vs 110±5 ng/mmol creatinine). The chromatographic profile indicates that pro-IGF-II may also be present. The concentrations of IGF-II appear to be less variable than the other reported parameters. The mean IGF binding protein-1 concentrations in these urine samples was 414±83 ng/mmol creatinine. IGFs in the urine are in part bound to binding proteins.

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