THE EFFECT OF DIFFERENT EXTRACTION PROCEDURES ON TWO DIFFERENT MOLECULAR WEIGHT SPECIES OF SERUM NSILA AND ON THE CARRIER PROTEIN OF SMALL MOLECULAR WEIGHT NSILA (NSILA-S)

1979 ◽  
Vol 90 (4) ◽  
pp. 637-647 ◽  
Author(s):  
U. Kaufmann ◽  
J. Zapf ◽  
E. R. Froesch
Keyword(s):  
Molecular Weight ◽  
Acetic Acid ◽  
Binding Protein ◽  
Extraction Procedure ◽  
Gel Filtration ◽  
Adsorption Chromatography ◽  
Carrier Protein ◽  
List Type ◽  
Small Molecular Weight ◽  
Binding Characteristics

ABSTRACT The influence of Dowex-50 adsorption chromatography on the recovery of two different forms of serum NSILA, large and small mol. wt. NSILA, and on the recovery of the binding protein of the small mol. wt. form was studied and compared with another extraction procedure, gel filtration on Sephadex G-50 in 1 m acetic acid. Partially purified NSILA-S is adsorbed to Dowex-50 at pH 6.8. It can be eluted with 20 mm NH4OH and appears unchanged with regard to its biological activity and molecular weight. Adsorption of 125I-labelled NSILA-S to Dowex-50 does not change its binding characteristics to serum. When serum is chromatographed on Sephadex G-50 in 1 m acetic acid, NSILA is obtained in a large and in a small molecular weight form (NSILA-S). After recombination of the small molecular weight NSILA fraction with the "stripped" serum fraction, which contains large mol. wt. NSILA and a specific carrier protein for NSILA-S, re-chromatography of this mixture on Sephadex G-50 at neutral pH yields NSILA mostly in the void volume. It adsorbs to Dowex-50. After elution from Dowex, acidic gel filtration on Sephadex G-50 results in an elution pattern which is completely different from that of NSILA-S. Adsorption of serum to Dowex-50 results in a dramatic decrease of the NSILA-S binding activity. It is concluded that Dowex-50 adsorption chromatography of serum inactivates most of the serum NSILA-S binding protein leads to the loss of acid dissociable small mol. wt. NSILA (NSILA-S). Therefore, Dowex-50 adsorption chromatography is not suitable for the subsequent determination or further purification of NSILA-S from whole serum.

Zinc- and copper-binding proteins in the plasma of winter flounder (Pseudopleuronectes americanus)

1980 ◽  
Vol 58 (4) ◽  
pp. 609-613 ◽  
Author(s):  
P. E. Fletcher ◽  
G. L. Fletcher
Keyword(s):  
Molecular Weight ◽  
Binding Proteins ◽  
Binding Protein ◽  
Gel Filtration ◽  
Protein S ◽  
Egg Yolk ◽  
Winter Flounder ◽  
Zinc Binding ◽  
Copper Binding ◽  
Copper Binding Proteins

Zinc- and copper-binding proteins were isolated from the plasma of winter flounder using gel filtration chromatography. A single copper-binding protein fraction of molecular weight 170 000 was isolated from the plasma of both sexes.In male and female flounder over 95% of the plasma zinc was associated with a zinc-binding protein(s) with a molecular weight of 76 000. In male flounder the remaining zinc appeared to be bound to a protein(s) of molecular weight 186 000. In female flounder the remaining 5% of the zinc was associated with two zinc-binding fractions with apparent molecular weights of 186 000 and 340 000 – 370 000.Extracts of plasma vitellogenin and egg yolk proteins revealed significant quantities of zinc and copper. It is hypothesized that the female specific zinc-binding protein (340 000 – 370 000) was vitellogenin.

Concentrations and Kinetics of Phosphorus Fractions Along the Trophic Gradient of Lake Memphremagog

1979 ◽  
Vol 36 (8) ◽  
pp. 970-979 ◽  
Author(s):  
Robert Henry Peters
Keyword(s):  
Molecular Weight ◽  
Total Phosphorus ◽  
Gel Filtration ◽  
Soluble Reactive Phosphorus ◽  
Small Molecular Weight ◽  
P Deficiency ◽  
Large Molecular Weight ◽  
Soluble P ◽  
Phosphorus Production ◽  
Total P

Published relationships between chlorophyll and total phosphorus concentrations show that the amount of chlorophyll developed per unit of total P increases as total P increases. If the availability of phosphorus changes with lake trophy, this should be reflected in the proportion of total P found in different analytical fractions. From June 1975 to September 1976, the concentrations of total phosphorus, soluble phosphorus, orthophosphate, large molecular weight phosphorus (> 5000 mol wt) and small molecular weight phosphorus (< 400 mol wt) were measured at six sites along the phosphorus-production gradient of Lake Memphremagog; 32P-PO4 was used to follow short-term exchanges between available orthophosphate and other fractions. Total P declined threefold among sites but the proportion of different fractions remained relatively constant which suggests that any disproportionate loss from one compartment is made good by exchanges with the others. On average, soluble P formed\of total P, soluble reactive phosphorus formed 14%, and PO4 8%; gel filtration showed that ~40% of soluble P was associated with large molecular weight phosphorus and ~40% with small molecular weight phosphorus. Within this basic similarity, was a trend to decreased PO4 concentrations and increased concentrations of soluble P and soluble unreactive phosphorus at more oligotrophic sites. Tracer experiments showed that all fractions exchanged some P with 32P-PO4, although soluble fractions, especially large molecular weight phosphorus, interacted more slowly with orthophosphate than did particulate P. Faster orthophosphate turnover suggested greater P deficiency in more oligotrophic sites. This deficiency may promote the formation of a refractory soluble P which may explain a part of the reduced ratio of chlorophyll to phosphorus when total P is reduced. Key words: orthophosphate, phosphorus, turnover, availability, lakes, gel fitration, soluble organic P

scholarly journals Studies on the appearance of a hepatic copper-binding protein in normal and zinc-deficient rats

1976 ◽  
Vol 36 (1) ◽  
pp. 101-112 ◽  
Author(s):  
I. Bremner ◽  
N. T. Davies
Keyword(s):  
Molecular Weight ◽  
Metal Binding ◽  
Binding Protein ◽  
Gel Filtration ◽  
Apparent Molecular Weight ◽  
Low Molecular Weight ◽  
Copper Binding ◽  
Hepatic Copper ◽  
Copper Binding Protein ◽  
Molecular Weight Form

1. A study has been made by gel-filtration techniques of the soluble copper- and zinc-binding proteins in rat liver after both intraperitoneal injection of Cu and dietary Cu supplementation.2. Liver Cu and Zn concentrations increased after injection of Cu, both metals accumulating in the cytosol, mainly in a fraction with an apparent molecular weight of (about 12 000)3. When Zn-deficient rats were injected with Cu, there was little change in liver Zn concentration and the occurrence of Cu in the low-molecular-weight form (about 12 000) was more transient. At most periods after injection, Cu accumulated mainly in a fraction with a molecular weight greater than 65 000.4. When the rats were Cu-loaded by dietary supplementation, virtually no Cu or Zn was found in the low-molecular-weight form in Zn-deficient rats, although they were found in the Zn-supplemented animals.5. The results suggest that Zn is essential for the accumulation of Cu in this form, but not for Cu to stimulate production of the metal-binding protein by a process requiring active protein synthesis.

scholarly journals The Use of Different Proteins as a Carrier Protein to Obtaining Morphine-Protein Conjugates for ELISA Diagnosis of Drug Addicts

2021 ◽  
pp. 296-303
Author(s):  
Farkhod Eshboev ◽  
Elvira Yusupova ◽  
Galina Piyakina ◽  
Sabirdjan Sasmakov ◽  
Jaloliddin Abdurakhmanov ◽  
...  
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Drug Addiction ◽  
Soybean Protein ◽  
Gel Filtration ◽  
Carrier Protein ◽  
Drug Addicts ◽  
Soybean Variety ◽  
Filtration Method ◽  
Protein Conjugates

Drug addiction is one of the biggest problems of medicine because diagnosis and treatment of drug addiction are difficult compared with some other socially significant diseases. In this study, synthesis and evaluation of four carrier protein-morphine conjugates were experimented. These conjugates were evaluated based on ELISA; soybean protein-based conjugate was selected for further analysis. The total soybean protein was isolated from the local soybean variety and; it was fractioned by the gel-filtration method and their amino acids compositions were studied. After that, the ELISA drug addicts were conducted based on soybean protein-morphine conjugates synthesized with soybean protein fractions. The high molecular weight soybean protein- morphine conjugate showed the highest quality.

BCL-2α encodes a novel small molecular weight GTP binding protein

1990 ◽  
Vol 30 ◽  
pp. 145-153 ◽  
Author(s):  
Subrata Haldar ◽  
Christine Beatty ◽  
Carlo M. Croce
Keyword(s):  
Molecular Weight ◽  
Binding Protein ◽  
Small Molecular Weight ◽  
Gtp Binding Protein ◽  
Gtp Binding

Bioactive parathyroid hormone in canine progressive renal insufficiency

1984 ◽  
Vol 247 (4) ◽  
pp. E442-E448
Author(s):  
D. Grunbaum ◽  
M. Wexler ◽  
M. Antos ◽  
M. Gascon-Barre ◽  
D. Goltzman
Keyword(s):  
Molecular Weight ◽  
Renal Failure ◽  
Parathyroid Hormone ◽  
Renal Insufficiency ◽  
Final Stage ◽  
Gel Filtration ◽  
Small Molecular Weight ◽  
Dihydroxyvitamin D ◽  
Severe Renal Disease ◽  
Circulating Levels

Bioactive parathyroid hormone and hormonal actions were monitored as hyperparathyroidism evolved in a model of progressive canine renal failure. Circulating levels of bioactive and immunoreactive parathyroid hormone rose as renal insufficiency worsened, but elevations, especially in bioactivity, were most marked in the final stage of uremia. By gel filtration analysis, the major circulating bioactive moiety was similar to the major glandular form of parathyroid hormone, although a smaller-molecular-weight entity was seen in the final stage of renal failure. Renal phosphate threshold fell, urinary hydroxyproline corrected for glomerular filtration rose, and plasma 1,25-dihydroxyvitamin D fell but remained detectable, as renal function deteriorated. The results demonstrate a progressive rise in bioactive parathyroid hormone, show the appearance of a small-molecular-weight bioactive entity in severe renal disease, and correlate effects of the rising bioactive parathyroid hormone with changes in renal phosphate handling and with skeletal resorption.

Fate of receptor-bound human chorionic gonadotrophin in pseudopregnant rat ovaries perfused in vitro

1985 ◽  
Vol 109 (1) ◽  
pp. 115-121
Author(s):  
Hannu Rajaniemi ◽  
Jan Sogn ◽  
Paul Holmes ◽  
Björn Källfelt ◽  
Per Olof Janson
Keyword(s):  
Molecular Weight ◽  
Histological Examination ◽  
Gel Filtration ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Cell Periphery ◽  
Small Molecular Weight ◽  
Immunohistochemical Studies

Abstract. Pseudopregnant rats were injected with [125I] hCG, anaesthesized 1 h later and after cannulation of the aorta the ovaries were isolated and perfused with Gey & Gey buffer containing 0.2% BSA. The release of radioactivity was monitored for 2 h and analyzed by gel filtration. Five to ten per cent of the radioactivity was released within 2 h and represented small molecular weight peptides and iodotyrosine and [125I]hCG. Analysis of the ovarian radioactivity prior to and after perfusion revealed that virtually all hCG was receptor-bound. Loading the medium with unlabelled hCG displaced [125I]hCG from the receptor but did not enhance its degradation. Histological examination showed that the ovarian tissues were still intact after the 2 h perfusion. Immunohistochemical studies revealed a localization of the hCG at the cell periphery both prior to and after perfusion. These results provide evidence showing that the rate of internalization of receptor-hCG complexes in rat luteal cells is slow in vivo.

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