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

Author(s):  
Farkhod Eshboev ◽  
Elvira Yusupova ◽  
Galina Piyakina ◽  
Sabirdjan Sasmakov ◽  
Jaloliddin Abdurakhmanov ◽  
...  

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.

1984 ◽  
Vol 62 (5) ◽  
pp. 276-279 ◽  
Author(s):  
C. H. Lin ◽  
W. Chung ◽  
K. P. Strickland ◽  
A. J. Hudson

An isozyme of S-adenosylmethionine synthetase has been purified to homogeneity by ammonium sulfate fractionation, DEAE-cellulose column chromatography, and gel filtration on a Sephadex G-200 column. The purified enzyme is very unstable and has a molecular weight of 120 000 consisting of two identical subunits. Amino acid analysis on the purified enzyme showed glycine, glutamate, and aspartate to be the most abundant and the aromatic amino acids to be the least abundant. It possesses tripolyphosphatase activity which can be stimulated five to six times by S-adenosylmethionine (20–40 μM). The findings support the conclusion that an enzyme-bound tripolyphosphate is an obligatory intermediate in the enzymatic synthesis of S-adenosylmethionine from ATP and methionine.


2021 ◽  
Vol 18 (1) ◽  
pp. 13-19
Author(s):  
A.I. Rabadanova ◽  

The steady growth in the number of drug addicts, especially among young people, dictates the need to find ways to prevent and treat this disease. In this regard, there is a need for a more detailed study of the mechanisms of the course of this disease using modern research methods, such as atomic force microscopy and fluorescence analysis of amino acid residues. Purpose of the work: to reveal the structural and functional state of erythrocyte membranes in drug addiction. Materials and methods. The studies were carried out on the erythrocyte membranes of 60 subjects suffering from heroin addiction. The shape and topography of the erythrocyte surface were studied, and spectral analysis of the proteins of the erythrocyte membranes was carried out. Results. The conducted AFM studies of erythrocyte membranes indicate the heterogeneity of the surface mechanical properties of the erythrocyte membranes of drug addicts. The data obtained indicate an acceleration of the aging process of erythrocytes in drug addiction, which goes in two ways: the formation of outgrowths on the plasmolemma, which subsequently die off (echinocytes) and invagination of the plasmolemma of erythrocytes (spherocytes). The fluorescence spectrum of amino acids in erythrocytes of drug addicts is characterized by a significant decrease in the intensity of almost all peaks and a shift of the fluorescence peak to the short-wave region. Findings. With drug addiction, changes in the structural integrity of red blood cells are noted. In people with drug addiction, in comparison with healthy people, there is a higher variability of the morphology of erythrocytes, which is expressed in a significant increase in the proportion of echinocytes and spherocytes against the background of a significant decrease in the number of discocytes. For the membrane proteins of erythrocytes of drug addicts, conformational changes are characteristic, manifested in a decrease in the intensity of fluorescence of aromatic amino acids, which indicates their structural modification and significant vulnerability of the hematopoietic system. They are largely determined by changes in the fluorescence intensity of tryptophan and, to a lesser extent, tyrosine, which indicates the preservation of the three-dimensional structure of the protein.


1981 ◽  
Author(s):  
H R Lijnen ◽  
B Wiman ◽  
B Van Hoef ◽  
D Collen

α2-Antiplasmin (α2AP), the main physiological inhibitor of plasmin in human plasma, is a single–chain glycoprotein with a molecular weight of 67,000 consisting of about 510 amino acids and containing 13 percent carbohydrate.A tryptic digest on 400 mg of reduced, carboxymethylated and citraconylated purified α2AP was performed. Peptides were separated by combinations of ion exchange chromatography, gel filtration and high performance liquid chromatography, and sequenced using the manual Edman degradation. Some peptides were further digested in order to establish overlaps. At the time of submission of this abstract we have sequenced 7 out of the approximately 21 arginyl peptides completely (each between 3 and 21 residues) and are working on the others. At present we have about 200 residues of sequence. Here we only report the stretches of 10 amino acids or more, which may be useful to compare the structure of α2AP with that of other serine protease inhibitors.


1977 ◽  
Vol 55 (24) ◽  
pp. 4257-4266 ◽  
Author(s):  
Lewis A. Slotin ◽  
Denis R. Lauren ◽  
Ross E. Williams

Several polypeptides have been synthesized which contain the alternating sequence lysyl-X, where X = gly, L-ala, D-ala, L-val, L-leu, and L-phe. The polypeptides have been characterized by gel filtration (molecular weight) and by circular dichroism spectroscopy (secondary structure).


1971 ◽  
Vol 122 (5) ◽  
pp. 623-631 ◽  
Author(s):  
Anne M. S. Marr ◽  
A. Neuberger ◽  
Wendy A. Ratcliffe

1. Tamm–Horsfall glycoprotein from rabbit urine has been isolated and characterized. The homogeneity of the preparation has been established by a variety of procedures including disc gel electrophoresis and ultracentrifugation in aqueous solution, sodium dodecyl sulphate and formic acid. 2. The chemical composition has been determined and a carbohydrate content of approx. 31% was obtained. The relative contents of the amino acids were shown to be very similar to those in human Tamm–Horsfall glycoprotein. A trace of lipid was also detected. 3. Leucine was identified as the only N-terminal amino acid. 4. The subunit structure was investigated in the presence of sodium dodecyl sulphate by gel filtration and disc gel electrophoresis. These studies indicated that the subunit possessed a molecular weight of approx. 84000±6000. A similar value was obtained after reduction and S-alkylation of the glycoprotein indicating that the disulphide bonds were all intrachain. 5. A minimum value for the chemical molecular weight of 85000±6000 was obtained from the number of N-terminal amino acids released by cyanogen bromide cleavage of the glycoprotein. 6. The immunological properties of the glycoprotein were studied. Cross reactivity was demonstrated between human Tamm–Horsfall glycoprotein and a guinea-pig anti-rabbit Tamm–Horsfall antiserum.


Soil Research ◽  
1968 ◽  
Vol 6 (2) ◽  
pp. 225 ◽  
Author(s):  
GD Swincer ◽  
JM Oades ◽  
DJ Greenland

After the removal of light fraction from soils under old pasture and under continuous fallow-wheat rotation, carbohydrates were extracted using IN HC1 followed by 0.5N NaOH and finally an acidic acetylation procedure, or by a single extraction with 0.2N NaOH only. The sequential extraction procedure removed 70-80 % of the carbohydrate from the soil under both agronomic systems. 0.2N NaOH removed a larger proportion of the carbohydrates from soil under fallow-wheat rotation (43-52%) than from soil under old pasture (35-38%). The composition of the carbohydrates in a given extract from the soil under pasture or fallow-wheat was similar. This similarity extended even to the neutral sugar composition of fractions obtained by gel filtration of the purified extracts. Generally, low molecular weight materials were rich in amino acids and compounds such as glucose, ribose, and glycerol. Polymers of molecular weight 4000-100,000 contained relatively high proportions of uronic acids and amino acids. Least amino acids were present in materials of molecular weight greater than 100,000 which contained appreciable quantities of deoxyhexoses (up to 20% of the total neutral sugars) indicative of their microbial origin. Against this background of similarity, certain differences between the carbohydrates from soils under pasture and fallow-wheat rotation were apparent. 1N HCl extracts contained more high molecular weight material from the old pasture soils than from the cultivated soil. The composition of these extracts indicated that they comprised the easily extractable recently synthesized microbial polysaccharides. The proportion of such polymers was lower in the cropped soil. A higher proportion of materials of small size was present in soils under a wheat crop. Maximum amounts of these compounds were present during periods of maximum plant and microbial activity. Extracts from soils under fallow-wheat rotation contained a higher proportion of uronic and amino acids and less ribose, arabinose, and deoxysugars than the extracts from soils under pasture. Based on relative deoxysugar contents it was calculated that the pasture soil contains about four times as much microbial polysaccharide as the soil under fallow-wheat.


1979 ◽  
Vol 42 (05) ◽  
pp. 1607-1614 ◽  
Author(s):  
A Diaz Batista ◽  
G Hernandez Solana ◽  
J F Corral Almonte

SummaryA plasminogen activating substance was purified from the dialysates of the eluates of glass adsorbed kallikrein from fresh human plasma, by chromatography on QAE-Sephadex A-50 and gel filtration in Sephadex G-25. The preparation was concentrated by lyophilization. Its electrophoretic mobility was found to be similar to that of prealbumin. Its molecular weight appeared to be 15000–18000 daltons. The analysis of aminoacids of this activator showed that it contains a high proportion of acid amino acids. The purified activator showed esterase activity, fibrinolytic activity and kininogenase activity on heated human plasma. These activities were respectively equivalent to 150 μM BAEe/mg protein, 19 × 103 units of streptokinase/μg protein and 250 μg bradykinin/mg protein.


1981 ◽  
Vol 199 (1) ◽  
pp. 9-15 ◽  
Author(s):  
M Janusz ◽  
K Starościk ◽  
M Zimecki ◽  
Z Wieczorek ◽  
J Lisowski

A proline-rich polypeptide isolated from sheep colostrum is described. The molecular weight of the polypeptide determined by gel filtration is 17 200. However, in the presence of guanidinium chloride the molecular weight found is about 6000. The polypeptide contains about 22% of proline, a high proportion of non-polar amino acids, a low percentage of glycine, and no alanine, arginine and cysteine residues. The only N-terminal amino acid found is leucine. C.d. spectra in water and in 50% (v/v) trifluoroethanol suggest the presence of block sequences of proline residues forming helices of polyproline II type. The proline-rich polypeptide is soluble at 4 degrees C but is reversibly precipitated on warming to room temperature. Maximal precipitation is observed at pH 4.6 and at ionic strength above 0.6. The precipitation depends on the concentration of the polypeptide. No effect of other proteins, Ca2+ and Zn2+ ions on the precipitation of the polypeptide was found. The proline-rich polypeptide is not an amphipathic protein. The lack of effect of the polypeptide on proteolytic enzymes ruled out the possibility that it is an inhibitor of proteinases.


1976 ◽  
Vol 56 (3) ◽  
pp. 159-166 ◽  
Author(s):  
G. GUIDI ◽  
G. PETRUZZELLI ◽  
P. SEQUI

The distribution of individual amino acids and monosaccharides in fulvic acid and its fractions separated by polyamide chromatography was investigated in five different Italian soils. Although little differences were generally found in the two polyamide fractions (FI and FII), the highest percentage content of acidic amino acids and the lowest percentage content of neutral amino acids have been found in the second one (FII); monosaccharides composition was more irregular, but generally FII contained more pentoses. Both chromatographic fractions (FI and FII) have been chromatographed on Sephadex G-25. The composition in carbohydrate and amino acid components of the further different fractions resolved by gel filtration showed great differences depending on the molecular weight distribution.


1979 ◽  
Vol 90 (4) ◽  
pp. 637-647 ◽  
Author(s):  
U. Kaufmann ◽  
J. Zapf ◽  
E. R. Froesch

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.


Sign in / Sign up

Export Citation Format

Share Document