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141.
Regulation of acetyl-CoA carboxylase in rat mammary gland. Effects of starvation and of insulin and prolactin deficiency on the fraction of the enzyme in the active form in vivo 总被引:12,自引:12,他引:0 下载免费PDF全文
The `initial' (I), endogenous phosphatase-activated (A) and citrate-activated (C) activities of acetyl-CoA carboxylase were measured in mammary-gland extracts of pregnant and lactating rats. There was a 10-fold increase in the A and C enzyme activities in the transition from early to peak lactation [cf. data of Mackall & Lane (1977) Biochem. J. 162, 635–642], but there was no significant increase in the ratio of the initial activity to the A and C activities of the enzyme. Starvation (24h) or short-term (3h) streptozotocin-induced diabetes both resulted in a 40% decrease in I/A and I/C activity ratios. In starvation this was accompanied by a decrease in the absolute values of the A and C activities such that the initial activity in mammary glands of starved animals was 45% that in glands from fed animals. Insulin treatment of starved or diabetic animals 60min before killing increased the I activity without affecting the A or C enzyme activities. Removal of the pups for 24h from animals in peak lactation (weaning) resulted in a marked but similar decrease in all three activities such that, although the initial activity was only 10% of that in suckled animals, the I/A and I/C activity ratios remained high and unaltered. Inhibition of prolactin secretion by injection of 2-bromo-α-ergocryptine gave qualitatively similar results to those during weaning. Simultaneous administration of ovine prolactin completely prevented the effects of bromoergocryptine. It is suggested that the initial activity of acetyl-CoA carboxylase in rat mammary gland is regulated by at least two parallel mechanisms: (i) an acute regulation of the proportion of the enzyme in the active state and (ii) a longer-term modulation of enzyme concentration in the gland. Insulin appeared to mediate its acute effects through mechanism (i), whereas prolactin had longer-term effects on enzyme concentration in the gland. A comparison of initial enzyme activities (I) obtained in the present study with rates of lipogenesis measured in vivo [Agius & Williamson (1980) Biochem. J. 192, 361–364; Munday & Williamson (1981) Biochem. J. 196, 831–837] gave good agreement between the two sets of data for all conditions studied except for 24h-starved and streptozotocin-diabetic animals. It is suggested that acetyl-CoA carboxylase activity is rate-limiting for lipogenesis in the mammary gland in normal, fed, suckled or weaned animals but that in starved and short-term diabetic animals changes in the activity of the enzyme by covalent modification alone may not be sufficient to maintain the enzyme in its rate-limiting role. 相似文献
142.
Victor J. Hruby 《Molecular and cellular biochemistry》1982,44(1):49-64
Summary Examination of glucagon structure-activity relationships and their use for the development of glucagon antagonists (inhibitors) have been hampered until recently by the lack of high purity of semisynthetic glucagon analogs and inadequate study of full dose-response curves for these analogs in sensitive bioassay systems. Recently a number of highly purified glucagon fragments and semi-synthetic analogs have been prepared and their full dose-response activities examined over a wide concentration range using the hepatic membrane adenylate cyclase assay, the hepatic membrane receptor binding assay, and glycogenolytic activity in isolated rat hepatocytes. The results of these studies have enabled us to identify and dissociate the structural (and in some cases conformational) features of glucagon important for binding from those most responsible for biological activity (transduction). Key findings in these studies were the observation that: (1) the C-terminal region of glucagon is primarily of importance for hormone binding to receptors; (2) glucagon1–21 and glucagon1–6 have low potency, but are essentially fully active glucagon derivatives; and (3) highly purified glucagon2–29 ([1-des-histidine]-glucagon), [1-N-carbamoylhistidine]-glucagon and [1-N-carbamoylhistidine, 12-N-carbamoyllysine]-glucagon are all partial agonists.These and other findings led us to synthesize several semisynthetic analogs of glucagon which were found to possess no intrinsic biological activity in the hepatic adenylate cyclase assay system, but which could block the effect of glucagon (competitive inhibitors) in activating adenylate cyclase in this system. Two of these highly purified analogs [1-des-histidine] [2-N-trinitrophenylserine, 12-homoarginine]-glucagon and [1-N-trinitrophenylhistidine, 12-homoarginine]-glucagon were quite potent glucagon antagonists (inhibitors) with pA2 values of 7.41 and 8.16 respectively. The latter compound has also been demonstrated to decrease dramatically blood glucose levels of diabetic animals in vivo. These results demonstrate that glucagon is a major contributor to the hyperglycemia of diabetic animals.Examination of the known and calculated conformational properties of glucagon provide insight into the structural and conformational properties of glucagon and its analogs most responsible for its biological activity. Consideration of these features and the mechanism of glucagon action at the membrane receptor level provide a framework for further developing glucagon analogs for theoretical and therapeutic applications. 相似文献
143.
144.
David F. Smith John L. Magnani Victor Ginsburg 《Archives of biochemistry and biophysics》1981,209(1):52-56
Proteins that bind carbohydrates can be used to isolate specific sugar sequences from complex mixtures. Free sialyloligosaccharides or sialyloligosaccharides released from gangliosides by ozonolysis and alkaline fragmentation are labeled at their reducing ends by reduction with NaB[3H]4. After partial separation by column chromatography, oligosaccharide fractions are tested for binding to anti-sialyloligosaccharide antibodies [Smith, D. F., and Ginsburg, V. (1980) J. Biol. Chem.255, 55–59] and cholera toxin by a nitrocellulose filter assay. Oligosaccharides bound by the proteins can be eluted from the filters and further characterized. The method can be used to isolate and identify carbohydrate ligands of cell surfaces. 相似文献
145.
Identification of a Cholinergic-Specific Antigen Chol-1 as a Ganglioside 总被引:15,自引:11,他引:4
Peter J. Richardson John H. Walker R. Theresa Jones Victor P. Whittaker 《Journal of neurochemistry》1982,38(6):1605-1614
Abstract: An antiserum specific for cholinergic terminals was used to identify an antigen conserved between Elasmobranchs and mammals. Immunohistochemistry and a cytotoxicity test were used to assay the binding of antibody to mammalian terminals. Torpedo electric organ gangliosides totally abolished antibody binding. The highest inhibitory activity was associated with a single polysialoganglioside band on TLC plates. Neuraminidase altered the migration of the inhibitory activity on TLC plates. Antibody binding was inhibited by ganglioside fractions derived from chicken and mammalian brains. A summary of those tissues in which the antigen has been detected is presented. The possible function of the antigen is discussed. 相似文献
146.
Postnatal Changes in the Activity Ratio of Specific and Nonspecific Cholinesterases from Neuronal Perikarya 总被引:3,自引:2,他引:1
Enrique L. M. Ochoa Alicia Brusco Victor P. Idoyaga-Vargas† Hector Carminatti† 《Journal of neurochemistry》1982,38(5):1236-1240
Abstract: A soluble fraction from rat brain neuronal perikarya was shown to contain both the specific and nonspecific forms of the enzyme acetylcholines-terase (EC's 3.1.1.7. and 3.1.1.8., respectively). The ratio of the enzyme activities varied along the course of brain development: the nonspecific form being predominant from 1 to 15 days of age and the specific one showing the pattern of rising activity from day 15 onward. We suggest a possible relationship between this changing in cholinesterase activities and the establishment of synapses within the rat cerebral cortex. 相似文献
147.
Analysis of Lepidium sativum seeds showed the presence of allyl, 2-phenethyl and benzyl glucosinolates, the first two being reported for the first time from this source. The effects of temperature, pH of the extraction medium and the length of time allowed for autolysis were assessed on the benzyl glucosinolate degradation products in seed extracts. In particulàr benzyl thiocyanate was not produced at higher temperatures but at ambient and lower temperatures it exceeded isothiocyanate. Nitrile was always the major product under the conditions studied, ever at pH levels as high as 7.4. Five new possible benzyl glucosinolate degradation products were detected and evidence is presented that benzaldehyde and benzyl alcohol could be secondary products formed thermally from isothocyanate and thiocyanate, respectively. Benzyl mercaptan and benzyl methyl sulphide also appear to be thermally produced. 相似文献
148.
Angie Rizzino Victor Terranova David Rohrbach Craig Crowley Heather Rizzino 《Journal of cellular biochemistry》1980,13(2):243-253
In this paper we have examined the growth and differentiation of the embryonal carcinoma cell line, F9, in the defined medium EM-3 at low density. We show that the growth of F9 and their differentiated cells (F9-diff) in EM-3 is strongly density dependent. At low cell densities the growth of both cell types is severely limited and most of the cells do not survive. Although this poses a problem for working with F9 and F9-diff in EM-3, it provides a convenient assay for identifying molecules that support their growth at low density. Using this assay, we have determined that laminin, a newly isolated glycoprotein of basement membranes, significantly improves the growth and short-term survival of both F9 and F9-diff. However, addition of laminin to EM-3 is insufficient to promote the clonal growth of these cell types. Our findings also indicate that laminin promotes the attachment of F9 and F9-diff in defined media. On the basis of our results, we propose an attachment function for laminin during the early stages of mammalian development. 相似文献
149.
Achiasmatic meiosis and complex heterozygosity in female cyclopoid copepods (Copepoda,Crustacea) 总被引:1,自引:0,他引:1
In Mesocyclops edax S.A. Forbes, 2n=14, a North American copepod, the females are heterozygous for several interchanges leading to the formation of large rings of chromosomes (rings of 14, or of 12 plus 1 bivalent) at meiotic metaphase, comparable to those of the plant Oenothera, although no chiasmata are present. The chromosomes are more or less metacentric and have large terminal H-segments. In the rings homologous arms are held together by connecting fibers which insert close to the euchromatin-heterochromatin junctions. Coordinated orientation of the zigzag type seems to be the role. 相似文献
150.
Victor H. Kollman John L. Hanners Robert E. London Enrique G. Adame Thomas E. Walker 《Carbohydrate research》1979,73(1):193-202
The blue-green alga Agmenellum quadruplicatum (strain PR6) has been used to prepare photobiosynthetically 13C-labeled d-glucose, 2-O-(α-d-glucopyranosyl)-glyceric acid (glucosylglycerate), 2-hydroxy-1-(hydroxymethyl)ethyl α-d-gluco-pyranoside (glucosylglycerol), and α-d-glueopyranosyl β-d-fructofuranoside (sucrose). When grown to a cell density of 4.4 g.L-1 (dry weight) under nitrate-nitrogen limiting growth conditions for 120 h, the algal cells contained 38% of the dry-cell weight as(1 → 4)-α-d-glucan (amylose). About 1% of the dry-cell weight was glucosylglycerol, glucosylglycerate, and sucrose. Glutamate was obtained, together with carbohydrates of low molecular weight, when the cells were extracted with chloroform-methanol; d-glucose was recovered from the extracted cells by acid hydrolysis of the starch. The algae were grown by using 20 mol% [13C] carbon dioxide for preparation of labeled carbohydrates and for cellular component identification by whole-cell n.m.r. spectroscopy. 相似文献