Purification and Some Properties of the Soluble and Membrane-Bound Adenosine Deaminases of Micrococcus sodonensis ATCC 11880 and their Distribution within the Family Micrococcaceae

1975 ◽  
Vol 53 (3) ◽  
pp. 344-353 ◽  
Author(s):  
M. A. Pickard
Keyword(s):  
Adenosine Deaminase ◽  
Gel Filtration ◽  
Membrane Vesicles ◽  
Soluble Enzyme ◽  
Adenosine Deaminases ◽  
Adenosine Deaminase Activity ◽  
The Family ◽  
Adenosine Transport ◽  
Membrane Bound ◽  
Isolated Membrane Vesicles

In Micrococcus sodonensis and some other Micrococcus species, adenosine deaminase is present both as a membrane-bound and a soluble enzyme. The membrane-bound adenosine deaminase can be extracted with n-butanol, and may account for up to 5% of the total cellular adenosine deaminase activity. In a number of comparative tests, no differences between the two enzyme forms could be found, thus they are believed to be similar molecular species. The purified membrane-bound or soluble enzyme had a molecular weight, obtained by gel-filtration, of 130 000 and was inactive toward adenine and adenine mononucleotides. It appears, therefore, to be more closely related to the calf-intestine enzyme than the Aspergillus oryzae form in respect to size and substrate specificity. Attempts to correlate membrane-bound adenosine deaminase activity with adenosine transport in isolated membrane vesicles of M. sodonensis indicated no obvious relationship between the two activities.

Metabolism and Transport of Purine Nucleosides by Membrane Preparations of Micrococcus sodonensis

1974 ◽  
Vol 52 (2) ◽  
pp. 83-89 ◽  
Author(s):  
M. A. Pickard ◽  
Lucille Phillippe ◽  
J. N. Campbell
Keyword(s):  
Adenosine Deaminase ◽  
Membrane Vesicles ◽  
Purine Nucleoside ◽  
Nucleoside Transport ◽  
Marked Contrast ◽  
Adenosine Concentration ◽  
Membrane Bound ◽  
Uptake And Metabolism ◽  
Isolated Membrane Vesicles ◽  
Bacterial Systems

The uptake and metabolism of adenosine-8-14C was studied using isolated membrane vesicles prepared from Micrococcus sodonensis. When the adenosine concentration was 1.0 mM, a concentration that caused prolonged bacteriostasis in growing cells, the membranes vesicles initially accumulated adenosine and inosine to similar levels but rapidly became enriched with inosine. However, when the external adenosine concentration was lowered to 0.1 mM, the vesicle contents were almost entirely inosine. No compounds other than adenosine and inosine were detected. These effects were shown to be the result of a membrane-bound adenosine deaminase and are in marked contrast to the phosphoribosyltransferase-linked mechanisms of purine nucleoside transport in other bacterial systems.

Passive diffusion of nucleosides into Micrococcus sodonensis membrane vesicles

1980 ◽  
Vol 58 (6) ◽  
pp. 457-460 ◽  
Author(s):  
M. A. Pickard
Keyword(s):  
Energy Source ◽  
Membrane Vesicles ◽  
Passive Diffusion ◽  
Membrane Bound ◽  
Nucleoside Uptake ◽  
Membrane Bound Enzymes ◽  
Isolated Membrane Vesicles

Nucleoside entry into isolated membrane vesicles of Micrococcus sodonensis (luteus) was studied using 14C-labelled nucleosides: adenosine, inosine, cytidine, uridine, guanosine, and thymidine. All nucleosides were recovered unmetabolized from the vesicles except adenosine and cytidine which were partly deaminated by membrane-bound enzymes. Vesicle preparations actively transported proline but no energy source was found capable of supporting concentrative nucleoside uptake. The entry of nucleosides into M. sodonensis vesicles was not saturable, nor was there competition between the nucleosides studied for entry. It was concluded that nucleoside entry into M. sodonensis vesicles occurs by passive diffusion.

scholarly journals Adenosine deaminase activity in blood of patients with stable angina pectoris.

1997 ◽  
Vol 44 (2) ◽  
pp. 359-361 ◽  
Author(s):  
M Kopff ◽  
I Zakrzewska ◽  
J Klem ◽  
J Kalinowska-Fuchs ◽  
M Strzelczyk
Keyword(s):  
Angina Pectoris ◽  
Blood Plasma ◽  
Adenosine Deaminase ◽  
Normal Level ◽  
Stable Angina ◽  
Control Group ◽  
Stable Angina Pectoris ◽  
Adenosine Deaminases ◽  
Adenosine Deaminase Activity

The activity of adenosine deaminases (EC.3.5.4.4) in granulocytes and lymphocytes of patients with stable angina pectoris was lower by about 27% and 24%, respectively as compared with control group, whereas these values in erythrocytes and blood plasma were at the normal level.

scholarly journals Preliminary characterization of two thymus-dependent xenoantigens from mouse lymphocytes

1977 ◽  
Vol 163 (2) ◽  
pp. 211-217 ◽  
Author(s):  
I S Trowbridge ◽  
M Nilsen-Hamilton ◽  
R T Hamilton ◽  
M J Bevan
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Membrane Vesicles ◽  
Sodium Deoxycholate ◽  
Pea Lectin ◽  
Preliminary Characterization ◽  
Low Concentrations ◽  
Membrane Bound

Preliminary characterization of two mouse thymus-dependent (T) lymphocyte xenoantigens, T25 and T200, which are selectively labelled by lactoperoxidase-catalysed iodination of T-cells, is described. Both molecules are membrane-bound glycoproteins. Fractionation of membrane vesicles prepared from BW5147 lymphoma cells by sedimentation through sucrose density gradients show that antigens T25 and T200 are in fractions enriched with plasma membrane. Moreover antigen T200 is partially degraded when viable cells are treated briefly with low concentrations of trypsin. Both molecules are efficiently solubilized in buffers containing sodium deoxycholate or Nonidet P-40, as measured by failure to sediment at 100000g for 60min. However, gel filtration on Sepharose 6B showed the presence of aggregated material in Nonidet P-40 extracts which was not found in deoxycholate-solubilized membranes. After solubilization in detergent, antigens T25 and T200 bind to, and may be specifically eluted from, columns of pea lectin--Sepharose or concanavalin A--Sepharose. Both molecules are heterogeneous when examined by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. As judged by its binding to columns of pea lectin, at least part of the heterogeneity of mouse thymocyte antigen T25 resides in its carbohydrate moiety.

Half-Life of Platelet Factor 4 (PF-4) in Plasma and Platelets from Macaca Mulatta

1977 ◽  
Vol 37 (01) ◽  
pp. 073-080 ◽  
Author(s):  
Knut Gjesdal ◽  
Duncan S. Pepper
Keyword(s):  
Macaca Mulatta ◽  
Half Life ◽  
Human Platelet ◽  
Gel Filtration ◽  
Platelet Factor 4 ◽  
Active Protein ◽  
Platelet Factor ◽  
Membrane Bound

SummaryHuman platelet factor 4 (PF-4) showed a reaction of complete identity with PF-4 from Macaca mulatta when tested against rabbit anti-human-PF-4. Such immunoglobulin was used for quantitative precipitation of in vivo labelled PF-4 in monkey serum. The results suggest that the active protein had an intra-platelet half-life of about 21 hours. In vitro 125I-labelled human PF-4 was injected intravenously into two monkeys and isolated by immuno-precipita-tion from platelet-poor plasma and from platelets disrupted after gel-filtration. Plasma PF-4 was found to have a half-life of 7 to 11 hours. Some of the labelled PF-4 was associated with platelets and this fraction had a rapid initial disappearance rate and a subsequent half-life close to that of plasma PF-4. The results are compatible with the hypothesis that granular PF-4 belongs to a separate compartment, whereas membrane-bound PF-4 and plasma PF-4 may interchange.

Rotavirus interaction with isolated membrane vesicles.

1994 ◽  
Vol 68 (6) ◽  
pp. 4009-4016 ◽  
Author(s):  
M C Ruiz ◽  
S R Alonso-Torre ◽  
A Charpilienne ◽  
M Vasseur ◽  
F Michelangeli ◽  
...  
Keyword(s):  
Membrane Vesicles ◽  
Isolated Membrane Vesicles

scholarly journals Adenosine deaminase activity in rheumatoid pleural effusion.

1988 ◽  
Vol 47 (5) ◽  
pp. 394-397 ◽  
Author(s):  
I Ocana ◽  
E Ribera ◽  
J M Martinez-Vazquez ◽  
I Ruiz ◽  
E Bejarano ◽  
...  
Keyword(s):  
Pleural Effusion ◽  
Adenosine Deaminase ◽  
Adenosine Deaminase Activity

scholarly journals Improving cell-free glycoprotein synthesis by characterizing and enriching native membrane vesicles

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jasmine M. Hershewe ◽  
Katherine F. Warfel ◽  
Shaelyn M. Iyer ◽  
Justin A. Peruzzi ◽  
Claretta J. Sullivan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Synthetic Biology ◽  
Membrane Protein ◽  
Membrane Vesicles ◽  
Processing Methods ◽  
Glycoprotein Synthesis ◽  
On Demand ◽  
Free Gene ◽  
Membrane Bound

AbstractCell-free gene expression (CFE) systems from crude cellular extracts have attracted much attention for biomanufacturing and synthetic biology. However, activating membrane-dependent functionality of cell-derived vesicles in bacterial CFE systems has been limited. Here, we address this limitation by characterizing native membrane vesicles in Escherichia coli-based CFE extracts and describing methods to enrich vesicles with heterologous, membrane-bound machinery. As a model, we focus on bacterial glycoengineering. We first use multiple, orthogonal techniques to characterize vesicles and show how extract processing methods can be used to increase concentrations of membrane vesicles in CFE systems. Then, we show that extracts enriched in vesicle number also display enhanced concentrations of heterologous membrane protein cargo. Finally, we apply our methods to enrich membrane-bound oligosaccharyltransferases and lipid-linked oligosaccharides for improving cell-free N-linked and O-linked glycoprotein synthesis. We anticipate that these methods will facilitate on-demand glycoprotein production and enable new CFE systems with membrane-associated activities.

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