Dopamine Accumulation in Papaver somniferum Latex

1984 ◽  
Vol 39 (11-12) ◽  
pp. 1034-1037 ◽  
Author(s):  
B. C. Homeyer ◽  
Margaret F. Roberts
Keyword(s):  
Papaver Somniferum ◽  
Dopamine Uptake ◽  
Chromaffin Granules ◽  
Catecholamine Uptake ◽  
Ph Dependent

Abstract Organelles found in the 1000 x g fraction of latex taken from Papaver somniferum have been shown to accumulate dopamine to the exclusion of other catecholamines. Dopamine accumula­ tion is both temperature and pH dependent and there is no evident requirement for ATP (1 mᴍ) or Mg2+ (2mᴍ). The relatively high (4.37 mᴍ), specificity of dopamine uptake, and insensitivity to inhibitors distinguish papaver latex organelle dopamine uptake from catecholamine uptake by mammalian chromaffin granules.

Enhancement by F-actin of MgATP-dependent dopamine uptake into isolated chromaffin granules

IUBMB Life ◽  
1996 ◽  
Vol 40 (1) ◽  
pp. 61-66
Author(s):  
Kyoji Morita ◽  
Stuart Tomares ◽  
Harvey Pollard
Keyword(s):  
Dopamine Uptake ◽  
Chromaffin Granules

scholarly journals The ionogenic nature of the secretory-granule membrane. Electrokinetic properties of isolated chromaffin granules

1972 ◽  
Vol 130 (3) ◽  
pp. 825-832 ◽  
Author(s):  
E. K. Matthews ◽  
R. J. Evans ◽  
P. M. Dean
Keyword(s):  
Surface Charge ◽  
Negative Charge ◽  
Single Type ◽  
Chromaffin Granules ◽  
Chromaffin Granule ◽  
Granule Membrane ◽  
Dose Dependent Fashion ◽  
Ph Dependent ◽  
Dose Dependent ◽  
Anionic Surface

1. Chromaffin granules isolated from the bovine adrenal medulla possess an electrophoretic mobility of -1.12μm·s-1·cm·V-1, corresponding to a surface ζ potential of -14.4mV and surface charge density of 1.38×10-6C·cm-2. 2. The mobility of chromaffin granules is pH-dependent, indicating an amphoteric surface with an isoelectric point at pH3.0 and acidic groups with a pKa of 3.11. 3. Addition of bi- and ter-valent cations decreased the mobility of chromaffin granules in a dose-dependent fashion with a relative potency of La3+»Mn2+>Ca2+ >Sr2+>Mg2+>Ba2+. 4. Treatment with neuraminidase decreased the mobility of erythrocytes by 84%, whereas chromaffin-granule mobility was decreased by only 14%. This correlates well with the small complement of neuraminic acid present in the granule membrane. 5. The nature, origin and significance of the anionic surface charge of the chromaffin granule is discussed. It is concluded that the net negative charge at the surface of shear derives chiefly from a single type of chemical group, namely -CO2-, contributed by the α-carboxyl group of constituent proteins, the phospholipid phosphatidylserine and, to a lesser extent, the sialic acid component of glycoproteins.

Catecholamine Uptake and Release in Isolated Chromaffin Granules Exposed to Halothane

1980 ◽  
Vol 53 (5) ◽  
pp. 385-389 ◽  
Author(s):  
Koji Sumikawa ◽  
Yoshikuni Amakata ◽  
Kiyoshi Yoshikawa ◽  
Takeshi Kashimoto ◽  
Futoshi Izumi
Keyword(s):  
Chromaffin Granules ◽  
Catecholamine Uptake ◽  
Uptake And Release

scholarly journals A characterization of the nucleotide uptake by chromaffin granules of bovine adrenal medulla

1978 ◽  
Vol 172 (3) ◽  
pp. 353-360 ◽  
Author(s):  
Werner Aberer ◽  
Herwig Kostron ◽  
Ewald Huber ◽  
Hans Winkler
Keyword(s):  
Adenosine Triphosphatase ◽  
Alkaline Ph ◽  
Chromaffin Granules ◽  
Temperature Dependent ◽  
Ph Optimum ◽  
Catecholamine Uptake ◽  
Electrical Component ◽  
Atp Analogues ◽  
Dependent Mechanism

Chromaffin granules isolated from bovine adrenal gland were incubated with 3H-labelled nucleotides and [14C]noradrenaline to study the uptake of these substances. [3H]ATP, [3H]ADP and [3H]AMP are taken up by these organelles by the same temperature-dependent mechanism. The apparent Km for ATP and ADP is 1.4mm, and for AMP it is 2.9mm. The uptake of ATP has a flat pH optimum, whereas the catecholamine uptake increases with more alkaline pH. Atractyloside and carboxyatractyloside are competitive and specific inhibitors of nucleotide uptake, whereas reserpine inhibits only that for catecholamines. Mg2+ ions activate uptake of both catecholamine and nucleotides, whereas EDTA and N-ethylmaleimide inhibit these processes. Nucleotide and catecholamine uptakes are inhibited by uncouplers of oxidative phosphorylation and by two ATP analogues. NH4+ ions and nigericin in the presence of KCl inhibit only catecholamine uptake. It is concluded that nucleotide uptake, as proposed previously for catecholamine uptake, depends on an electrochemical proton gradient produced by a proton-translocating adenosine triphosphatase localized in the membrane of chromaffin granules. Furthermore, as suggested by the effect of NH4+ and nigericin, catecholamine uptake apparently depends on the chemical part of this gradient, whereas the results for nucleotide uptake are consistent with its dependence on the electrical component.

Immunogold labeling of the calcium binding protein synexin in isolated adrenal chromaffin granules and chromaffin cells

1990 ◽  
Vol 48 (3) ◽  
pp. 952-953
Author(s):  
Gemma A.J. Kuijpers ◽  
Harvey B. Pollard
Keyword(s):  
Adrenal Medulla ◽  
Calcium Binding ◽  
Chromaffin Cells ◽  
Cell Activation ◽  
Structural Information ◽  
Dependent Manner ◽  
Chromaffin Granules ◽  
Immuno Electron Microscopy ◽  
Ultrathin Sections ◽  
Adrenal Chromaffin

Exocytotic fusion of granules in the adrenal medulla chromaffin cell is triggered by a rise in the concentration of cytosolic Ca2+ upon cell activation. The protein synexin, annexin VII, was originally found in the adrenal medulla and has been shown to cause aggregation and to support fusion of chromaffin granules in a Ca2+-dependent manner. We have previously suggested that synexin may there fore play a role in the exocytotic fusion process. In order to obtain more structural information on synexin, we performed immuno-electron microscopy on frozen ultrathin sections of both isolated chromaffin granules and chromaffin cells.Chromaffin granules were isolated from bovine adrenal medulla, and synexin was isolated from bovine lung. Granules were incubated in the presence or absence of synexin (24 μg per mg granule protein) and Ca2+ (1 mM), which induces maximal granule aggregation, in 0.3M sucrose-40m MMES buffer(pH 6.0). Granules were pelleted, washed twice in buffer without synexin and fixed with 2% glutaraldehyde- 2% para formaldehyde in 0.1 M phosphate buffer (GA/PFA) for 30 min. Chromaffin cells were isolated and cultured for 3-5 days, and washed and incubated in Krebs solution with or without 20 uM nicotine. Cells were fixed 90 sec after on set of stimulation with GA/PFA for 30 min. Fixed granule or cell pellets were washed, infiltrated with 2.3 M sucrose in PBS, mounted and frozen in liquid N2.

The Reaction of Thiol Compounds with the Vitamin-K Dependent Clotting Factors

1969 ◽  
Vol 21 (03) ◽  
pp. 573-579 ◽  
Author(s):  
P Fantl
Keyword(s):  
Prothrombin Time ◽  
Vitamin K ◽  
Anaerobic Conditions ◽  
Clotting Factors ◽  
Thiol Compounds ◽  
Aerobic Conditions ◽  
One Stage ◽  
Factor Ii ◽  
Ph Dependent ◽  
The One

SummaryTreatment of human and dog oxalated plasma with 0.2 to 1.0 × 10−1 M 2.3-dithiopropanol (BAL) or dithiothreitol (DTT) at 2–4° C for 30 min results in the reduction of the vitamin-K dependent clotting factors II, VII, IX and X to the respective-SH derivatives. The reaction is pH dependent. Under aerobic conditions the delayed one stage prothrombin time can be partly reversed. Under anaerobic conditions a gradual prolongation of the one stage prothrombin time occurs without reversal.In very diluted plasma treated with the dithiols, prothrombin can be converted into thrombin if serum as source of active factors VII and X is added. In contrast SH factors VII, IX and X are inactive in the specific tests. Reoxidation to active factors II, VII, IX and X takes place during adsorption and elution of the SH derivatives. The experiments have indicated that not only factor II but also factors VII, IX and X have active-S-S-centres.

COMBINING ABILITY FOR INHERITANCE OF ECONOMIC TRAITS IN OPIUM POPPY (Papaver somniferum L.)

2011 ◽  
Vol 24 (2) ◽  
pp. 01-08
Author(s):  
B. Kumar ◽  
N. K. Patra
Keyword(s):  
Combining Ability ◽  
Diallel Cross ◽  
Papaver Somniferum ◽  
Opium Poppy ◽  
Economic Traits ◽  
Degree Of Dominance ◽  
Parental Material ◽  
The Mean ◽  
Per Se ◽  
Important Medicinal Plant

Opium poppy (Papaver somniferum L.) is an important medicinal plant produces more than 80 alkaloids obtained from the capsules and straw of the plant. The estimate of combining ability gives an indication of the genetic behaviour of the parental material. It is therefore, desirable to select the parents for hybridization on the basis of their per se performance and combining ability effects. The F1, F2 and their reciprocals of an eight parent diallel cross in opium poppy were studied for combining ability of seven economic traits. The mean sum of squares due to GCA, SCA and reciprocals were significant for all the traits. The magnitude of GCA variances were invariably higher than those of SCA and thus indicating the preponderance of non-additive genetic variances, which was further affirmed by the measure of average degree of dominance i.e. ?(?2s/? 2g).  Among the parents VN35I for plant height, Sanchita and VG26 for capsules per plant, VN23, VN35I and Vivek for capsule index, VG26 and Sanchita for seed and straw yield per plant, and VN35I and VG20 for morphine content were found good general combiners. Earliness being a desired trait, parent VG20 having significant negative GCA estimates coupled with per se performance can be considered as good general combiner for early flowering. Inclusion of good general combiners in a multiple crossing program or an inter-mating population involving all possible crosses among them subjected to bi-parental mating may be expected to offer maximum promise in breeding for economic traits.DOI: http://dx.doi.org/10.3329/bjpbg.v24i2.17000

Rationalizing the pH-Activity Response for Escherichia Coli’s Glycinamide Ribonucleotidetransformylase Through Computational Methods

2019 ◽  
Author(s):  
Adrian Roitberg ◽  
Pancham Lal Gupta
Keyword(s):  
Transfer Reaction ◽  
Catalytic Mechanism ◽  
Ph Dependence ◽  
Ph Effects ◽  
Dependent Manner ◽  
E Coli ◽  
Gar Tfase ◽  
Activity Curve ◽  
Ph Dependent ◽  
Formyl Transfer

<div>Human Glycinamide ribonucleotide transformylase (GAR Tfase), a regulatory enzyme in the de novo purine biosynthesis pathway, has been established as an anti-cancer target. GAR Tfase catalyzes the formyl transfer reaction from the folate cofactor to the GAR ligand. In the present work, we study E. coli GAR Tfase, which has high sequence similarity with the human GAR Tfase with most functional residues conserved. E. coli GAR Tfase exhibits structural changes and the binding of ligands that varies with pH which leads to change the rate of the formyl transfer reaction in a pH-dependent manner. Thus, the inclusion of pH becomes essential for the study of its catalytic mechanism. Experimentally, the pH-dependence of the kinetic parameter kcat is measured to evaluate the pH-range of enzymatic activity. However, insufficient information about residues governing the pH-effects on the catalytic activity leads to ambiguous assignments of the general acid and base catalysts and consequently its catalytic mechanism. In the present work, we use pH-replica exchange molecular dynamics (pH-REMD) simulations to study the effects of pH on E. coli GAR Tfase enzyme. We identify the titratable residues governing the pH-dependent conformational changes in the system. Furthermore, we filter out the protonation states which are essential in maintaining the structural integrity, keeping the ligands bound and assisting the catalysis. We reproduce the experimental pH-activity curve by computing the population of key protonation states. Moreover, we provide a detailed description of residues governing the acidic and basic limbs of the pH-activity curve.</div>

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