Journal of cyclic nucleotide research最新文献

Journal of cyclic nucleotide research Pub Date : 2006-01-01 DOI: 10.1038/mp.a001693.01

D. Communi, B. Robaye, J. Boeynaems

引用次数: 1

Partial purification and characterization of particulate guanylate cyclase from rat liver after solubilization with trypsin. 胰蛋白酶溶解大鼠肝脏中颗粒鸟苷酸环化酶的部分纯化和特性研究。

Journal of cyclic nucleotide research Pub Date : 1982-01-01

S A Waldman, J A Lewicki, H J Brandwein, F Murad

{"title":"Partial purification and characterization of particulate guanylate cyclase from rat liver after solubilization with trypsin.","authors":"S A Waldman, J A Lewicki, H J Brandwein, F Murad","doi":"","DOIUrl":"","url":null,"abstract":"Guanylate cyclase from 105,000 X g particulate fractions of rat liver homogenates (20 pmoles of cyclic GMP formed/min/mg protein) was solubilized in the absence of detergents by incubating fractions 12 min at 37 degrees with 5 ug/ml trypsin. Optimal solubilization was dependent upon trypsin and particulate preparation concentrations. Virtually no activation of particulate guanylate cyclase was observed at any time point or trypsin concentration tested. Guanylate cyclase solubilized with trypsin was purified about 500-fold (9.4 nmoles/min/mg protein) using ammonium sulfate precipitation, GTP-affinity chromatography, and preparative polyacrylamide gel electrophoresis. Activity eluted as a single peak on Sepadex G-200 (Stokes radius = 40 A) and migrated as a single peak on sucrose density gradients (S20,w = 4.6). Thus, the tryptic fragment was estimated to be about 80,000 daltons (Mr) with a frictional ratio (f/fo) of 1.4. These partially purified preparations exhibited linear double reciprocal plots with Mn-GTP and Hill coefficients of 1.0. This is in contrast to the crude membrane-associated enzyme which had a Hill co-efficient of 1.5. Membrane-bound and trypsin-solubilized guanylate cyclase were activated 3- and 4-fold with nitric oxide and were inhibited with 1mM cystamine. Cystamine inhibition could be partially reversed with 7.5 mM dithiothreitol. These studies indicate that particulate guanylate cyclase solubilized by limited proteolysis is amenable to purification by \"classical\" chromatographic techniques. The partially purified fragment contains the catalytic site, the site for nitric oxide activation, and at least one sulfhydryl group required for activity.","PeriodicalId":15497,"journal":{"name":"Journal of cyclic nucleotide research","volume":"8 6","pages":"359-70"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17202607","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Rabbit kidney cortex phosphorylase phosphatases: evidence for complexes between high molecular weight forms and heat-stable inhibitor proteins. 兔肾皮质磷酸化酶磷酸酶:高分子量形式和热稳定抑制剂蛋白之间复合物的证据。

Journal of cyclic nucleotide research Pub Date : 1982-01-01

R L Mellgren, K K Schlender

{"title":"Rabbit kidney cortex phosphorylase phosphatases: evidence for complexes between high molecular weight forms and heat-stable inhibitor proteins.","authors":"R L Mellgren, K K Schlender","doi":"","DOIUrl":"","url":null,"abstract":"Two forms of high molecular weight phosphorylase phosphatase have been partially resolved by gel filtration chromatography of rabbit kidney cortex extracts. Two heat-stable inhibitor proteins co-eluted with the phosphatase peaks. Phosphorylase phosphatase and heat-stable inhibitor activity also co-migrated on gel electrophoresis of cortex extracts. When extracts were heated to 95 degrees for 5 minutes prior to gel filtration or electrophoresis, phosphorylase phosphatase inhibitor activity eluted at a lower molecular weight and a higher mobility, respectively. Storing cortex extracts at -20 degrees for 6 months resulted in partial conversion of both phosphatase and inhibitor activities to lower molecular weight forms which co-eluted on gel filtration. The two inhibitor peaks from gel filtration chromatography were heat-treated and characterized. Both inhibitor peaks had molecular weight of 25,000 to 35,000. The inhibitory activity of one of the peaks was increased about 3.5-fold by incubation with cyclic AMP-dependent protein kinase and ATP, and required higher concentrations of TCA to be precipitated. Hence, one of the inhibitor peaks resembled rabbit muscle inhibitor -1, while the other peak may represent an inhibitor similar to rabbit muscle inhibitor -2. These results represent the first indication that low molecular weight heat-stable inhibitor proteins may be bound to high molecular weight phosphorylase phosphatases in the cell.","PeriodicalId":15497,"journal":{"name":"Journal of cyclic nucleotide research","volume":"8 1","pages":"27-37"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17352364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Functional compartments in cyclic nucleotide action. 环核苷酸作用的功能区室。

Journal of cyclic nucleotide research Pub Date : 1982-01-01

J S Hayes, L L Brunton

{"title":"Functional compartments in cyclic nucleotide action.","authors":"J S Hayes, L L Brunton","doi":"","DOIUrl":"","url":null,"abstract":"Cyclic AMP-dependent protein kinase (cAMP-PK) is a ubiquitous enzyme that, when activated by cAMP, is capable of phosphorylating a variety of intracellular proteins. The central postulate of cAMP-mediated hormone action is that hormones regulate intracellular cAMP concentration and cAMP-PK mediates the effects of this second messenger. Although this postulate accurately describes cAMP action in certain systems, it does not adequately provide for recent observations of the accumulation of cAMP and the activation of protein kinase without the anticipated effects on protein kinase's substrates. Both biochemical and cytochemical technics provide evidence that hormonally-specific regulation of cAMP action occurs and is important. Our thesis is that hormonal regulation of metabolic events via cAMP is localized intracellular phenomenon. We propose that occupation of some cell-surface hormone receptors leads to cAMP accumulation and the activation of protein kinase in subcellular compartments, with the consequent phosphorylation of specific, rather than all, substrates of protein kinase. circumstances potentially contributing to this specificity include: (a) physical and kinetic compartmentation of hormone-receptor-adenylate cyclase complexes non-randomly within the cell membrane; and, (b) a fixed spatial relationship of hormonally activated adenylate cyclase and specific intracellular regions by the participation of cytoskeletal proteins.","PeriodicalId":15497,"journal":{"name":"Journal of cyclic nucleotide research","volume":"8 1","pages":"1-16"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17352363","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Evidence for cross-linking of cyclic AMP to constituents of brain tissue by aldehyde fixatives: potential utility in histochemical procedures. 乙醛固定剂使环AMP与脑组织成分交联的证据:在组织化学过程中的潜在效用。

Journal of cyclic nucleotide research Pub Date : 1982-01-01

T W Rall, R A Lehne

{"title":"Evidence for cross-linking of cyclic AMP to constituents of brain tissue by aldehyde fixatives: potential utility in histochemical procedures.","authors":"T W Rall, R A Lehne","doi":"","DOIUrl":"","url":null,"abstract":"The amount of cyclic AMP recovered from unfixed tissue sections of brain carried through immunohistochemical procedures was found to be small (less than 2 pmoles/mg protein) and constant despite wide variations in the amount accumulated prior to freezing and sectioning. However, treatment of brain slices with glutaraldehyde or formaldehyde in buffered sucrose solutions for one to two hours at 0 degrees rendered insoluble as much as 60% of the total accumulated cyclic AMP as judged by filtration of homogenates of treated tissue. Immunoreactive cyclic AMP was recovered from filters by extraction with warm, dilute acid. The fraction of filter-bound cyclic AMP was relatively constant over a wide range of initial tissue levels. The persistence of insoluble cyclic AMP was greatest in homogenates of slices treated with formaldehyde when maintained above pH 8.5; about 40% of this fraction was lost after two hours at 0 degrees. Incubation of tissue homogenates with formaldehyde and radioactive nucleoside or nucleotide derivatives of adenine, guanine, and cytosine resulted in a time-dependent appearance of insoluble radioactivity; compounds lacking an amino function were inactive. Pure proteins, including polylysine, also reacted with formaldehyde and 3H-cyclic AMP to produce radioactivity resistant to adsorption by charcoal. The reaction between cyclic AMP or adenosine, formaldehyde, and alkyl amines was examined using UV spectrometry. It is tentatively concluded that at 0 degrees formaldehyde is capable of rapidly producing a methylene-bridged adduct between primary alkylamines and the N6-amino function of purine nucleosides or nucleotides. Application of these results to the development of immunohistochemical procedures for the cellular localization of cyclic AMP will require the generation of antibody preparations with high reactivity for cyclic AMP derivatized at the N6-position.","PeriodicalId":15497,"journal":{"name":"Journal of cyclic nucleotide research","volume":"8 4","pages":"243-65"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17362443","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The guanine nucleotide-binding regulatory component of adenylate cyclase in human erythrocytes. 人红细胞中腺苷酸环化酶的鸟嘌呤核苷酸结合调控成分。

Journal of cyclic nucleotide research Pub Date : 1982-01-01

E Hanski, A G Gilman

引用次数: 0

Inhibition of 3',5'-cyclic nucleotide 3'-phosphohydrolase by a novel 1',2'-cyclic nucleotide. 新型1′，2′环核苷酸对3′，5′-环核苷酸3′-磷酸水解酶的抑制作用。

Journal of cyclic nucleotide research Pub Date : 1982-01-01

V Nair, R J Wiechert, D A Young

引用次数: 0

Enhancement of depolarization-dependent neurosecretion from PC12 cells by forskolin-induced elevation of cyclic AMP. 福斯克林诱导的环AMP升高对PC12细胞去极化依赖性神经分泌的影响。

Journal of cyclic nucleotide research Pub Date : 1982-01-01

C S Rabe, J Schneider, R McGee

{"title":"Enhancement of depolarization-dependent neurosecretion from PC12 cells by forskolin-induced elevation of cyclic AMP.","authors":"C S Rabe, J Schneider, R McGee","doi":"","DOIUrl":"","url":null,"abstract":"The effects of elevated intracellular cyclic AMP on the release of neurotransmitters was studied using the clonal pheochromocytoma cell line, PC12, and forskolin, a direct activator of adenylate cyclase. Intracellular cyclic AMP concentrations ranging from 8 to 400 times basal levels were achieved with 0.1 to 100 uM forskolin. Unstimulated release of neurotransmitters was unchanged by any concentration of forskolin. However, K+-stimulated release of both norepinephrine (NE) and acetylcholine was enhanced by 0.1 to 10 uM forskolin. Release of NE elicited by depolarization with carbachol and veratridine also was enhanced by 1 uM forskolin. Enhancement of release was reversed by higher concentrations of forskolin, especially in the presence of a phosphodiesterase inhibitor (RO 20-1724) which caused very large increases in cyclic AMP content. The enhancement of transmitter release from the PC12 cells occurred without concomitant changes in agonist-stimulated ion flux through the acetylcholine receptor ion channel, or in depolarization-dependent uptake of 45Ca++. Thus, increasing the cyclic AMP content of PC12 cells fails to initiate neurosecretion but appears to facilitate some element in the secretion process subsequent to Ca++ influx.","PeriodicalId":15497,"journal":{"name":"Journal of cyclic nucleotide research","volume":"8 6","pages":"371-84"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17202608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The role of cyclic AMP in the regulation of exocytosis in the rat parotid gland: evidence obtained with the isoproterenol analog PI-39. 环AMP在大鼠腮腺胞吐调节中的作用:异丙肾上腺素类似物PI-39获得的证据。

Journal of cyclic nucleotide research Pub Date : 1982-01-01

T N Spearman, J P Durham, F R Butcher

{"title":"The role of cyclic AMP in the regulation of exocytosis in the rat parotid gland: evidence obtained with the isoproterenol analog PI-39.","authors":"T N Spearman, J P Durham, F R Butcher","doi":"","DOIUrl":"","url":null,"abstract":"The isoproterenol analog, PI-39, induced a dose-dependent release of alpha-amylase from rat parotid minces in vitro. This effect was blocked by propranolol, a beta-adrenergic antagonist. PI-39 alone had no effect on parotid cyclic AMP levels or on protein kinase activation as assessed by the activity ratios method. However, PI-39 produced a dose-dependent increase in these parameters when a phosphodiesterase inhibitor was added to tissue minces simultaneously with the isoproterenol analog. Both isoproterenol and PI-39 altered the phosphorylation state of at least three specific endogenous phosphoproteins in (32P)-Pi prelabelled minces. The presence of a phosphodiesterase inhibitor was not required to demonstrate the effects of PI-39 on protein phosphorylation. Studies of endogenous protein phosphorylation in parotid broken cell preparations demonstrated that the phosphorylation of at least two of the PI-39 and isoproterenol-affected phosphoproteins are influenced by cyclic AMP. This study demonstrates that, under certain conditions, it is possible to activate a cyclic AMP-regulated biological process without elevating the total cellular cyclic AMP concentration or the protein kinase activity ratio.","PeriodicalId":15497,"journal":{"name":"Journal of cyclic nucleotide research","volume":"8 4","pages":"225-34"},"PeriodicalIF":0.0,"publicationDate":"1982-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17252820","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Identification and characterization of endogenous inhibitors of polyamine-responsive protein kinase activity. 内源性多胺反应蛋白激酶活性抑制剂的鉴定和表征。

Journal of cyclic nucleotide research Pub Date : 1982-01-01

Y Morishita, A Sahai, C Akogyeram, V Hollis, T Oka, W Criss

引用次数: 0