共查询到20条相似文献,搜索用时 15 毫秒
1.
Immunoreactive ACTH levels were determined in whole plasma, eluate fractions of plasma and perfusates of anterior lobe extracts subjected to chromatography on Sephadex G-50 fine. In the fetal plasma, ACTH levels were higher on day 19 than on days 17, 18, 20 and 21. After birth, ACTH concentrations dropped to reach the lowest values in one week old newborns; thereafter they increased until weaning on day 21. They were then similar to those of non pregnant adult females. Three peaks of immunoreactive ACTH were present in all the chromatograms; the first one eluted near the void volume ("big" ACTH, PM approximately 44,000), the third one coeluted with human ACTH (1-39) ("little" ACTH, PM approximately 4,500) and the second one eluted midway between the 2 previous peaks ("intermediate" ACTH, PM approximately 13,000). During the last days of pregnancy, the proportion of the "little" form of ACTH in the fetal plasma showed a gradual increase whereas that of the "big" one decreased. On days 17, 19 and 21 of gestation the anterior lobes of fetal pituitary glands released in vitro these 3 forms of immunoreactive ACTH in the same proportions as those observed in the anterior lobes. In contrast, the proportions of the circulating forms of ACTH were quite different; the "little" one gradually increased and the "big" one decreased as gestation progressed. In vitro controlled tryptic digestion of the isolated "big" form led to the appearance of "intermediate" and "little" forms suggesting some transformations in the circulation of the ACTH forms released by the fetal hypophysis in vivo. 相似文献
2.
Clarke BL 《Biochemical and biophysical research communications》1999,266(2):542-546
The effect of incubation temperature and ligand competition was tested for (125)I-ACTH binding to isolated rat lymphocytes. AlphaMSH but not Agouti-like peptide was an effective competitive inhibitor for cell surface binding at 4 degrees C. Cells incubated with (125)I-ACTH at 37 degrees C rapidly associated ligand for 10 min and then gradually lost the radioactivity with time. Cells incubated with (125)I-ACTH at 4 degrees C accumulated ligand to only about half the maximal amount when compared to cells incubated at 37 degrees C for 10 min. Temperatures below 20 degrees C and toxins that block lysosomal degradation blocked the loss of cell-associated radioactivity. These results suggest the lymphocyte ACTH receptor is the Melanocortin 5 receptor and the receptor is internalized by endocytosis to deliver ligand to the lysosome. 相似文献
3.
We performed the following experiments to evaluate the accuracy of our newly developed radioimmunoassay for somatostatin: (1) Recovery of synthetic somatostatin added to human, porcine, and canine plasma with or without extraction with 67% acetone or 76% ethanol, using 3 different region-specific antibodies and, where applicable, 125I-labelled Tyr-1- or Tyr-11-substituted somatostatin or 125I-N-Tyr-somatostatin as tracers. The recovery of somatostatin corrected for losses inherent in the extraction procedure was close to 100%, and independent of species, antibody and tracer. Somatostatin 1-28 was extracted slightly less efficiently. Unextracted plasma interfered massively in the assay. (2) Pharmacokinetic experiments with infusion of somatostatin into 14 pigs and determination of metabolic clearance rate (MCR) and T-1/2. MCR was 27-38 ml/kg per min, independent of infusion rate (6.1 or 13 pmol/kg per min), extraction procedure or tracer. T-1/2 was 1.9 min. The infused somatostatin was not measurable in unextracted plasma. (3) Characterization of endogenous and exogenous, labelled and unlabelled somatostatin 1-14 in human plasma, using Sephadex G-50 columns at pH 7.5 and 9.0. Human plasma showed excess immunoreactivity eluting at the void volume whereas synthetic somatostatin was recovered quantitatively at the position of marker somatostatin when added to the plasma. The immunoreactivity of the tracers was decreased (125I-Tyr-11-somatostatin) or abolished (125I-N-Tyr- or 125I-Tyr-1-somatostatin) after incubation with plasma or void volume fractions of plasma subjected to gel filtration. Extracted plasma did not contain void volume immunoreactivity, but like whole plasma, small amounts of components which coeluted with intact somatostatin. 相似文献
4.
The direct extra-adrenal actions of adrenocorticotropin 1-39 (ACTH) on electrical (E) and mechanical (M) characteristics of canine atrial tissues (AT) were investigated in in vitro experiments. One hundred twenty-five mU/ml of ACTH 1-39 significantly augmented the catecholamine induced positive inotropism as seen by shortening the time to peak tension (10.6%, p = 0.01) and increasing peak isometric tension (3.5 times, p = 0.001). Effects on the M responses were inhibited by propranolol (10(-6) M) (P). ACTH did not significantly modify action potential E or M parameters during cholinergic receptor antagonism or alpha-adrenergic receptor antagonism. Existence of a specific ACTH receptor was demonstrated using 125I radioiodinated ACTH 1-24. Significant binding of 125I-ACTH to AT was observed. Intracellular C-AMP levels were also measured in AT using radioimmunoassay. Tissues were exposed to 125mU/ml ACTH 1-39 plus combinations of norepinephrine (10(-6) M) (NE) and P. ACTH alone did not elevate intracellular C-AMP levels. NE increased C-AMP levels were not further increased by ACTH. Exposure to antagonist returned elevated C-AMP levels to control values. In conclusion (1) ACTH augments the NE induced M positive inotropism of the beta adrenergic receptor system. (2) ACTH specifically binds to AT and (3) ACTH does not utilize the C-AMP second messenger system. 相似文献
5.
HCG receptors of porcine ovarian plasma membranes were loaded with 125I-hCG and covalently crosslinked with glutaraldehyde. The plasma membranes were labeled with tritium. The hCG receptor complex was solubilized in 1% SDS and subsequently chromatographed on Biogel P 300. 40-50% of 125I-radioactivity eluted near the void volume. 3H radioactivity in these fractions indicated the presence of crosslinked hormone receptor complexes. A second minor peak of 125I-radioactivity appeared at 55000 d. A third peak represented subunits of 125I-hCG. Analysis of the Biogel P 300 purified 125I-hCG receptor complex by polyacrylamide gel electrophoresis in sodium dodecyl sulfate resulted in two radioactivity peaks. The main peak corresponded to 90 000-100 000 d, the second peak was calculated to approximately 140 000 d. As a result the majority of the hormone receptor complex has a molecular weight of 90 000-100 000 d. Existence of subunits is discussed. 相似文献
6.
After intravenous injection of 125I-ACTH1-24 into rats, the highest concentration of 125I was found in kidneys, adrenal and liver. Addition of 5- and 30-fold excess unlabelled ACTH reduced the uptake of 125I by 50 and 68%, respectively, indicating that the adrenal uptake was specific. Pretreatment with dexamethasone decreased the adrenal uptake of 125I and caused adrenal atrophy. Chronic ACTH treatment increased the size of the adrenals, but did not affect the adrenal uptake of 125I. These experiments demonstrate selective uptake of 125I by the adrenals after administration of 125I-ACTH1-24. 相似文献
7.
A study was made of the enzymic degradation of (125)I-labelled parathyroid hormone by rat kidney microsomes. Incubation with microsomes resulted in rapid destruction of the labelled hormone. The microsomal factor was not separable by dialysis, and the reaction was favoured by pH values in the physiological range. Velocity of the reaction varied directly as the substrate concentration, and additional crude parathyroid hormone (trichloroacetic acid-precipitated, 3.68mg./ml.) inhibited destruction of labelled hormone. There was much less inhibition with added trichloroacetic acid-precipitated calcitonin (3.92mg./ml.) and virtually none with added pig insulin (3.80mg./ml.). Gel filtration of control medium on P6 (Bio-Gel) yielded one radioactive peak at the void volume. After incubation with microsomes three further peaks were obtained on gel filtration. Only the void-volume peak contained intact (125)I-labelled parathyroid hormone, indicating that the microsomal enzyme degraded labelled hormone to a number of smaller fragments. 相似文献
8.
Y Matuk 《Canadian journal of physiology and pharmacology》1983,61(11):1391-1395
In view of the electron microscopic evidence that silver does not penetrate cellular barriers, the distribution of radioactive silver in rat blood and subcellular fractions of liver, kidneys, spleen, and forebrain was studied. It was found that 24 h after a single intraperitoneal injection high levels of radioactivity were reached which decreased at different rates in the various tissues studied. In plasma, liver, and kidneys there was an initial rapid loss of radioactivity which was followed by a slower rate of loss. In the blood, forebrain, and spleen the loss of radioactivity was linear and somewhat slower than in the other three tissues. The cytosols of the liver and kidneys contained 60% while those of the forebrain and spleen contained 30% of the total radioactivity found in the tissue homogenates. Gel filtration on Sephadex G-75 showed that all cytosols contained two peaks of radioactivity; a high molecular weight peak which eluted just after the void volume and a low molecular weight peak. The amount of radioactivity in both peaks was, however, much lower in the chromatographic peaks of the forebrain and spleen than that found in those of the liver and kidneys. Furthermore, the spleen had a comparatively very small low molecular weight radioactive peak. In vitro experiments with liver cytosol showed similar results to those found in vivo in that the high molecular weight radioactive peak could be removed by heat. It is concluded that silver does enter cells and that silver thionein exists in the cytosols of forebrain, spleen, kidney, and liver. 相似文献
9.
Presence of ACTH and its receptor on a B lymphocytic cell line: a possible autocrine function for a neuroendocrine hormone 总被引:2,自引:0,他引:2
K L Bost E M Smith L B Wear J E Blalock 《Journal of biological regulators and homeostatic agents》1987,1(1):23-27
A mouse B lymphocytic cell line, designated BCL1, was found to produce immunoreactive ACTH and to secrete this molecule into culture supernates. The BCL1-derived ACTH induced Y-1 adrenal cells to undergo a steroidogeneic response and was eluted from gel filtration columns at a molecular weight similar to that expected for pituitary-derived ACTH. Furthermore, ACTH receptors were detected on the surface of BCL1 cells using indirect immunofluorescence and 125I-ACTH binding studies. Scatchard analysis demonstrated the presence of high and low affinity binding sites with dissociation constants of 4.5 x 10(-12) M and 2.8 x 10(-9) M, respectively. The production of both ACTH and its receptor by this B lymphocyte cell line suggests that an autocrine mechanism might be important for the maintenance of the transformed phenotype. 相似文献
10.
Uptake of iodinated human kidney alpha-D-mannosidase by rat liver- Association with membrane elements and stability in vivo and in vitro. 总被引:2,自引:2,他引:0
下载免费PDF全文
![点击此处可从《The Biochemical journal》网站下载免费的PDF全文](/ch/ext_images/free.gif)
D V Marinkovic S L Petrovic J V Martinovic J N Marinkovic 《The Biochemical journal》1976,159(3):729-736
1. Human kidney alpha-D-mannosidase (form A) was labelled with 125I to a specific radio-activity of approx. 2250muCi/mg of protein, essentially without loss of enzymic activity. The enzymic activity and radioactivity of the iodinated material also co-migrated in gel filtration and gel electrophoresis. 2. The binding of 125I-labelled mannosidase in vitro to particulate material in liver and kidney homogenates was of the other of 2 pg/mg of particulate material in liver and kidney homogenates was of the order of 2pg/mg of particulate protein withing 16h at 37 degrees C, and essentially zero in intervals of up to 60 min. The degradation in vitro of labelled exogenous mannosidase was of the order of 10-20pg/ 16th per mg of protein in postnuclear supernatant, and it was saturated entirely within 1h at 37 degrees C. 3. The binding of labelled mannosidase in vivo to particulate elements of liver homogenates 60 min after intravenous injection was at least 10 times higher in terms of specific radioactivity than the highest value attainable in vitro. Virtually all exogenous enzyme bound to liver particulate material could be recovered in macromolecular form after disruption of membranes by detergents. 4. The radioactive enzyme bound to liver particulate material could be detached almost completely by shearing, repeated freezing and thawing, and exposure to strong detergents under conditions that do not eliminate rough-endoplasmic-membrane structure. It could bot be released, however, by high salt concentration (0.5M-KC1) or by exposure to weak detergents such as Tween 80. The particle-bound enzyme should thus be associated with plasma membranes and lysosome-like elements. 5. Of the rat tissues studied, only liver could approach, within 60 min after the injection, the concentration of exogenous mannosidase found in the blood serum. The activity per g tissue weight fell progressively from liver (60% of serum value) to kidney (16% of serum value), lung (8% of serum vlaue), spleen (6% of serum value) and brain (0.9% of serum value). Most of the radioactive enzyme found in tissues other than liver appeared to be present in a free form, whereas in liver more than 50% of the labelled enzyme was associated with membrane elements. 相似文献
11.
The subcellular distribution of 125I-labelled HDL taken up by rat hepatocytes in vivo and in vitro has been studied with subcellular fractionation techniques: differential centrifugation and isopycnic centrifugation in sucrose gradients. 125I-labelled HDL bind to plasma membranes both in vivo and in vitro and part of the membrane-bound 125I-labelled HDL can be dissociated by the addition of unlabelled HDL. The hepatocytes also internalize 125I-labelled HDL. The 125I-labelled HDL accumulate, however, at different intracellular sites in the in vivo and in vitro situation. The subcellular distribution pattern of 125I-labelled HDL taken up by the cells in vivo is similar to that of the lysosomal marker enzyme acid phosphatase. Peak activity was found at a density of 1.20 g/ml. In vitro 125I-labelled HDL accumulate in an organelle with a medium density of about 1.13 g/ml. This distribution was similar to that of the plasma membrane marker 5'-nucleotidase. The subcellular distribution of radioactivity taken up in vivo was changed to lower density by incubating the cells with chloroquine, a drug known to render the lysosomes more boyant. Chloroquine had no effect on the distribution of 125I-labelled HDL taken up by hepatocytes in vitro. 相似文献
12.
Heparin catalyses the inhibition of two key enzymes of blood coagulation, namely Factor Xa and thrombin, by enhancing the antiproteinase activities of plasma antithrombin III and heparin cofactor II. In addition, heparin can directly inhibit the activation of Factor X and prothrombin. The contributions of each of these effects to the anticoagulant activity of heparin have not been delineated. We therefore performed experiments to assess how each of these effects of heparin contributes to its anticoagulant activity by comparing the effects of heparin, pentosan polysulphate and D-Phe-Pro-Arg-CH2Cl on the intrinsic pathway of coagulation. Unlike heparin, pentosan polysulphate catalyses only the inhibition of thrombin by plasma. D-Phe-Pro-Arg-CH2Cl is rapid enough an inhibitor of thrombin so that when added to plasma no complexes of thrombin with its inhibitors are formed, whether or not the plasma also contains heparin. Heparin (0.66 microgram/ml) and pentosan polysulphate (6.6 micrograms/ml) completely inhibited the intrinsic-pathway activation of 125I-prothrombin to 125I-prothrombin fragment 1 + 2 and 125I-thrombin. On the addition of thrombin, a good Factor V activator, to the plasma before each sulphated polysaccharide, the inhibition of prothrombin activation was demonstrable only in the presence of higher concentrations of the sulphated polysaccharide. D-Phe-Pro-Arg-CH2Cl also completely inhibited the intrinsic-pathway activation of prothrombin in normal plasma. The inhibitory effect of D-Phe-Pro-Arg-CH2Cl was reversed if thrombin was added to the plasma before D-Phe-Pro-Arg-CH2Cl. The inhibition of the activation of prothrombin by the three agents was also abolished with longer times with re-added Ca2+. Reversal of the inhibitory effects of heparin and pentosan polysulphate was associated with the accelerated formation of 125I-thrombin-antithrombin III and 125I-thrombin-heparin cofactor complexes respectively. These results suggest that the anticoagulant effects of heparin and pentosan polysulphate are mediated primarily by their ability to inhibit the thrombin-dependent activation of Factor V, thereby inhibiting the formation of prothrombinase complex, the physiological activator of prothrombin. 相似文献
13.
V E Ghazarossian R R Dent K Otsu M Ross B Cox A Goldstein 《Analytical biochemistry》1980,102(1):80-89
β-Endorphin-like peptides in blood plasma of normal human subjects were studied by means of a radioimmunoassay (RIA) and gel filtration. Plasma was extracted with silica gel, which was washed with water and 1 n HCl, and eluted with 50% acetone. Plasma extracts thus obtained and standard synthetic human β-endorphin yielded parallel RIA curves. Total immunoreactivity in normal donors ranged from 1.2 to 10.4 fmol/ml (21 subjects). The immunoreactivity was completely destroyed by treatment with papain. Gel filtration indicated the presence of three components-one of unknown nature at the void volume and the others at elution positions characteristic of β-lipotropin and β-endorphin. Recoveries of human β-endorphin and β-lipotropin added to plasma were 53 and 58%, respectively. Addition of N-ethylmaleimide to plasma or of aprotinin to blood immmediately following collection had no effect on the amount of total immunoreactivity. Furthermore, a large amount of β-endorphin-like immunoreactivity. The above results lead us to conclude that a β-endorphin-like immunoreactive peptide occurs naturally in plasma of normal human subjects. 相似文献
14.
Comparison of leucine enkephalin and adrenocorticotrophin effects on adrenal function in fetal and adult sheep 总被引:1,自引:0,他引:1
At Day 120-125 of gestation equimolar amounts of ACTH and leu-enkephalin injected in vivo provoked similar rises in plasma cortisol concentrations in chronically catheterized fetuses. There was no concomitant change in plasma DHEA concentrations, or in maternal cortisol concentrations. At term (Days 135-140) 2 out of 5 animals responded similarly to both leu-enkephalin and ACTH injections with a rise in plasma cortisol concentrations, but the other 3 animals, in which basal cortisol concentrations had already risen, showed no response to either agonist. In adult sheep, ACTH provoked a significant increase in the plasma cortisol concentrations, but equimolar amounts of leu-enkephalin were without effect. There was a significant output of cortisol in response to ACTH administration by collagenase-dispersed adrenal cells from term sheep fetuses in vitro. Leu-enkephalin had no effect on cortisol output from dispersed adrenal cells when added by itself, or with ACTH. We conclude that leu-enkephalin is able to function as a stimulator of pituitary-adrenal function during fetal life. The lack of effect of leu-enkephalin on adrenal cells implies that its action is exerted not directly at the adrenal gland, but indirectly at the level of the hypothalamus or pituitary through stimulation of the release of other corticotrophic substances. 相似文献
15.
Various intracellular organelles as well as outer cell membranes of bovine corpora lutea intrinsically contain gonadotropin receptors (Rao et al., J biol chem 256 (1981) 2628 [5]). In order to investigate whether exogenously added human choriogonadotropin (hCG) can internalize and bind to the intracellular sites, bovine luteal slices that had been carefully checked with respect to structural and functional integrity were incubated with 0.1 nM 125I-hCG. Following incubation, specific radioactivity was found to be associated with various intracellular organelles, but not with cytosol. The order of radioactivity uptake by subcellular organelles following a 2-h incubation was: Golgi medium greater than Golgi heavy greater than Golgi light greater than plasma membranes = rough endoplasmic reticulum greater than mitochondria-lysosomes- greater than nuclei. The 5'-nucleotidase activity and electron microscopic examination of the fractions revealed that the presence of radioactivity in the intracellular organelles cannot be attributed solely to plasma membrane contamination. The internalization and intracellular binding of 125I-hCG was time and temperature-dependent. Only excess unlabeled hCG and hLH (but not hCG subunits, FSH and PRL) competed with 125I-hCG for internalization in luteal slices. Very little or no 125I-hCG added was internalized in liver or kidney slices; luteal, liver and kidney slices accumulated neither 125I-BSA nor 125I. The radioactivity eluted from various luteal subcellular organelles was able to rebind to fresh corresponding organelles and came off Sepharose 6B columns in a position corresponding to native 125I-hCG. The gel filtration profile of detergent-solubilized radioactivity revealed that 125I-hCG was macromolecular bound. The degraded and altered 125I-hCG was found in the incubation media. 相似文献
16.
Steroid metabolism in Nagase Analbuminemia Rats (NAR), a mutant strain established from Sprague-Dawley rats, was studied. NAR are characterized by lack of serum albumin and hyperlipidemia. Total testosterone concentration in the serum of NAR was lower than that of normal rats, while the serum free testosterone, LH and FSH concentrations were similar. The half lives of 14C-labeled testosterone administered intravenously in NAR and normal Sprague-Dawley (SD) rats were 4.4 and 4.1 min, respectively. The plasma clearance rates of testosterone in NAR and normal rats were 34.7 and 39.1 ml/min per kg body weight. On Sephadex G-100 chromatography, a mixture of [3H]testosterone and normal rat serum gave two protein peaks eluted in the void volume and the albumin fraction, and the radioactivity was eluted all in the albumin fraction. In contrast, a mixture of [3H]testosterone and NAR serum gave a single protein peak eluted in the void volume and the radioactivity was mainly eluted with this protein peak. The association constants of testosterone to NAR and normal rat sera were 1.25 and 2.24 X 10(4) M-1. Enzyme activities related to the synthesis of testosterone by the testicular microsomal fractions of NAR and normal rats were examined. The activities of 3 beta-hydroxysteroid dehydrogenase, 5-ene-4-ene isomerase, 17 alpha-hydroxylase, C-17-C-20 lyase and 17 beta-hydroxysteroid dehydrogenase were lower in NAR than in normal rats. The activity for synthesis of testosterone from pregnenolone by the testicular microsomal fraction of NAR was about 40% of that of normal rats. These findings indicate that the low serum concentration of testosterone in NAR is mainly attributable to decreased biosynthesis of testosterone in the testes. 相似文献
17.
Antithrombin activity of fucoidan. The interaction of fucoidan with heparin cofactor II, antithrombin III, and thrombin 总被引:6,自引:0,他引:6
F C Church J B Meade R E Treanor H C Whinna 《The Journal of biological chemistry》1989,264(6):3618-3623
Fucoidan, poly(L-fucopyranose) linked primarily alpha 1----2 with either a C3- or a C4-sulfate, is an effective anticoagulant in vitro and in vivo (Springer, G. F., Wurzel, H. A., McNeal, G. M., Jr., Ansell, N. J., and Doughty, M. F. (1957) Proc. Soc. Exp. Biol. Med. 94, 404-409). We have determined the antithrombin effects of fucoidan on the glycosaminoglycan-binding plasma proteinase inhibitors antithrombin III and heparin cofactor II. Fucoidan enhances the heparin cofactor II-thrombin reaction more than 3500-fold. The apparent second-order rate constant of thrombin inhibition by heparin cofactor II increases from 4 x 10(4) (in the absence of fucoidan) to 1.5 x 10(8) M-1 min-1 as the fucoidan concentration increases from 0.1 to 10 micrograms/ml and then decreases as fucoidan is increased above 10 micrograms/ml. The fucoidan reaction with heparin cofactor II-thrombin is kinetically equivalent to a "template model." Apparent fucoidan-heparin cofactor II and fucoidan-thrombin dissociation constants are 370 and 1 nM, respectively. The enhancement of thrombin inhibition by fucoidan, like heparin and dermatan sulfate, is eliminated by selective chemical modification of lysyl residues either of heparin cofactor II or of thrombin. The fucoidan-antithrombin III reactions with thrombin and factor Xa are accelerated maximally 285- and 35-fold at fucoidan concentrations of 30 and 500 micrograms/ml, respectively. Using human plasma and 125I-labeled thrombin in an ex vivo system, the heparin cofactor II-thrombin complex is formed preferentially over the antithrombin III-thrombin complex in the presence of 10 micrograms/ml fucoidan. Our results indicate that heparin cofactor II is activated by fucoidan in vitro and in an ex vivo plasma system and suggest that the major antithrombin activity of fucoidan in vivo is mediated by heparin cofactor II and not by antithrombin III. 相似文献
18.
In vivo metabolism of brain natriuretic peptide in the rat involves endopeptidase 24.11 and angiotensin converting enzyme 总被引:1,自引:0,他引:1
Y Vanneste S Pauwels L Lambotte M Deschodt-Lanckman 《Biochemical and biophysical research communications》1990,173(1):265-271
The metabolism of brain natriuretic peptide (BNP) was studied in rats infused with 125I-BNP. During the infusion, the intact peptide was progressively converted to labelled degradative products, separated into nine peaks of radioactivity on HPLC, and accounting for approximately 70% of total plasma radioactivity at the plateau phase. After stopping the infusion, intact BNP disappeared with a half-life of 1.23 +/- 0.35 min whereas the labelled fragments accounted for progressively greater proportion of total activity. The degradation of BNP was significantly reduced by phosphoramidon (t1/2, 11.28 +/- 0.49 min) and captopril (t1/2, 6.99 +/- 0.34 min). A maximal effect was observed when both protease inhibitors were given simultaneously (t1/2, 15.3 +/- 0.48 min). When 125I-BNP was incubated in vitro with purified endopeptidase 24.11 (E-24.11) and angiotensin-converting enzyme (ACE), there was a time-dependent disappearance of the intact peptide associated with the generation of six labelled fragments, corresponding to fragments found in vivo. In serum the peptide was rapidly degraded with a half-life of 4.6 +/- 0.1 min, and the pattern of labelled fragments was similar to that observed during in vitro incubation with ACE. Captopril significantly reduced the rate of degradation of BNP in serum. The results allow to associate two define enzyme activities, namely E-24.11 and ACE, with the metabolism of BNP in vitro. They also indicate that, despite a close homology between ANP and BNP, the two peptides undergo different pathways of clearance. 相似文献
19.
An in vitro heparin release of lipoprotein lipase (LPL) from whole blood, mainly from monocytes, was demonstrated by (1) the time-course of lipolytic activity with the presence of 10 U/ml heparin at 37 degrees C, (2) the distribution of LPL activity in monocyte and lymphocyte fractions, (3) an immuno-inactivation with anti-LPL immunoglobulin (IgG) and (4) responses to various compounds such as NaCl, protamine sulfate, heparin, and serum activator. The in vitro heparin-releasable LPL activity from blood correlated well with the LPL activity of postheparin plasma obtained from both normolipidemic and hyperlipidemic rabbits. Studies in humans revealed sex- and age-related variations in the in vitro heparin-releasable LPL from monocytes in the blood of 134 normal subjects and 24 hypertriglyceridemic subjects: The mean LPL activity was significantly higher in normal females over the age of 30, than in the corresponding males. In the hypertriglyceridemic group, the LPL activity was also higher in females than in males, but it was not significant. The in vitro heparin-releasable LPL activity from monocytes in blood was comparable to the LPL activity derived from adipose tissue and postheparin plasma, and thus it reflects lipoprotein metabolism. 相似文献
20.
A study was conducted to investigate whether human plasma contains one or more than one protein for the transport of vitamin D and of 25-hydroxyvitamin D (25-OH-D).Serum was labeled in vivo with a mixture of radioactive vitamin D3 (derived from orally administered tracer vitamin D3) and of endogenously synthesized labeled 25-OH-D3. Samples of such serum were subjected to several different protein fractionation procedures. Only a single peak of protein-bound radioactivity was observed after each of these procedures. The fraction comprising the ascending and the descending limbs of the single peak of protein-bound radioactivity (after each procedure) were separately pooled. In each instance the ratio of radioactive 25-OH-D3 to radioactive vitamin D3 was found to be almost identical in both the ascending and the descending limbs. Taken together, these findings provide strong evidence that human serum contains only a single binding protein responsible for the normal transport of both vitamin D and 25-OH-D. Plasma labeled in vitro with added 3H-labeled 25-OH-D3 was subjected to gel filtration on Sephadex G-200 and to chromatography on columns of DEAE-cellulose and of SP-Sephadex. After each of these procedures a single peak of protein-bound radioactivity was observed, with elution profiles of protein and of radioactivity that were identical with those observed with in vivo labeled serum. These data indicate that tracer 25-OH-D3 added to plasma in vitro binds to the same plasma protein normally responsible for the transport of vitamin D and of 25-OH-D. 相似文献