Evidence for the release at low salt concentration of a lipid–protein–carbohydrate complex from isolated envelopes and whole cells of a marine pseudomonad

1971 ◽  
Vol 17 (5) ◽  
pp. 713-723
Author(s):  
F. L. A. Buckmire ◽  
Robert A. MacLeod
Keyword(s):  
Salt Concentration ◽  
Analytical Ultracentrifugation ◽  
Divalent Cations ◽  
Sedimentation Coefficient ◽  
Whole Cells ◽  
Carbohydrate Complex ◽  
Low Salt ◽  
Cell Envelopes ◽  
Isolated Cell

The effect of divalent cations on the turbidity and pH of suspensions of isolated cell envelopes of a marine pseudomonad has been examined. As the concentration of Mg2+, Ca2+, or Mn2+ was increased the turbidity and pH of the suspensions increased. A concentration of 0.010 M of divalent cations produced the same turbidity in a suspension of cell envelopes as 1.0 M NaCl. The envelopes released soluble non-dialyzable material (Ndf) when suspended in 0.01 M NaCl but not when suspended in 0.001 M solutions of the divalent cations. Whole cells of the marine pseudomonad released an Ndf when suspended in 0.5 M sucrose. The Ndf from both cells and envelopes contained lipid, protein, and carbohydrate. Analytical ultracentrifugation of the Ndf from both cells and envelopes revealed the presence of only one major component with a sedimentation coefficient of 9.7 ± 0.07. Electrophoresis of Ndf gave rise to a component which stained for carbohydrate and another which stained for protein. Both moved to the anode.

Synthesis and conformation of sequential basic polypeptides with branched and linear side chains

1985 ◽  
Vol 50 (12) ◽  
pp. 2925-2936 ◽  
Author(s):  
Štěpánka Štokrová ◽  
Jan Pospíšek ◽  
Jaroslav Šponar ◽  
Karel Bláha
Keyword(s):  
Amino Acid ◽  
Salt Concentration ◽  
Salt Solution ◽  
Theoretical Work ◽  
Side Chain ◽  
Amino Acid Residues ◽  
Cd Spectra ◽  
Low Salt ◽  
Conformational Freedom ◽  
Basic Polypeptides

Polypeptides (Lys-X-Ala)n and (Lys-X-Gly)n in which X represents residues of isoleucine and norleucine, respectively, and polypeptide (Tle-Lys-Ala)n, were synthesized via polymerization of 1-hydroxysuccinimidyl esters of the appropriate tripeptides to complete previously studied series. Circular dichroism (CD) spectra of the respective polymers were measured as a function of pH and salt concentration of the medium. The results were correlated with those obtained previously with the same series containing different amino acid residues at the X-position. The helix forming ability of the polypeptides (Lys-X-Ala)n with linear X side chain was found to be independent of the length. In the series (Lys-X-Gly)n the unordered conformation was the most probable one except (Lys-Ile-Gly)n. This polymer assumed the β conformation even in low salt solution at neutral pH. An agreement with some theoretical work concerned with the restriction of conformational freedom of amino acid residue branching at Cβ atom with our experimental results is evident.

scholarly journals N 1 N 8-bis(γ-glutamyl)spermidine cross-linking in epidermal-cell envelopes. Comparison of cross-link levels in normal and psoriatic cell envelopes

1990 ◽  
Vol 271 (2) ◽  
pp. 305-308 ◽  
Author(s):  
N Martinet ◽  
S Beninati ◽  
T P Nigra ◽  
J E Folk
Keyword(s):  
Epidermal Cell ◽  
Skin Disease ◽  
Envelope Protein ◽  
Cell Envelope ◽  
Human Epidermis ◽  
Cross Linking ◽  
Cross Link ◽  
Gamma Glutamyl ◽  
Cell Envelopes ◽  
Isolated Cell

N1N8-Bis(gamma-glutamyl)spermidine was found in exhaustive proteolytic digests of isolated cell envelopes from human epidermis at levels comparable with those of epsilon-(gamma-glutamyl)lysine. Significantly higher than normal amounts of these compounds, particularly the bis(gamma-glutamyl)polyamine, were observed in envelopes from afflicted areas (scales) of psoriatic patients. These findings support the notions that N1N8-bis(gamma-glutamyl)spermidine, like epsilon-(gamma-glutamyl)lysine, functions in cell envelopes as an enzyme-generated protein cross-link and stabilizing force and that individuals with the chronic, recurrent skin disease, psoriasis, exhibit in involved epidermis abnormal cell-envelope-protein cross-linking.

Effect of Salt Concentration and Incubation Temperature on Formation of Histamine, Phenethylamine, Tryptamine and Tyramine During Miso Fermentation

1986 ◽  
Vol 49 (6) ◽  
pp. 423-427 ◽  
Author(s):  
K.-D. HENRY CHIN ◽  
P. E. KOEHLER
Keyword(s):  
Salt Concentration ◽  
Incubation Temperature ◽  
High Salt ◽  
Soybean Paste ◽  
Salt Level ◽  
Low Salt ◽  
Two Factors

Two factors, salt concentration and incubation temperature, were examined for their effect on the formation of histamine, phenethylamine, tryptamine and tyramine during miso (soybean paste) fermentation. Misos containing 5 and 10% NaCl were prepared and incubated at 25 and 35°C. The effect of each factor was determined from the chemical and microbiological changes in the misos during fermentation. Salt level was a significant factor in the formation of amines. Higher amine levels were found in low-salt (5% NaCl) formulations than in high-salt (10% NaCl) misos. Incubation temperature within the range of 25 to 35°C during fermentation had little effect on amine formation in misos.

scholarly journals THE CELL ENVELOPES OF TWO EXTREMELY HALOPHILIC BACTERIA

1963 ◽  
Vol 18 (3) ◽  
pp. 681-689 ◽  
Author(s):  
A. D. Brown ◽  
C. D. Shorey
Keyword(s):  
Single Unit ◽  
Cell Envelope ◽  
Halophilic Bacteria ◽  
Layered Compound ◽  
Halobacterium Halobium ◽  
Chemical Analyses ◽  
Unit Membrane ◽  
Thin Sections ◽  
Whole Cells ◽  
Cell Envelopes

The cell envelope of Halobacterium halobium was seen in thin sections of permanganate-fixed cells to consist of one membrane. This membrane appeared mostly as a unit membrane but in a few preparations it resembled a 5-layered compound membrane. The cell envelope of Halobacterium salinarium at high resolution was always seen as a 5-layered structure different in appearance from the apparent compound membrane of H. halobium. The "envelopes" which were isolated in 12.5 per cent NaCl from each organism were indistinguishable from each other in the electron microscope and comprised, in each case, a single unit membrane with an over-all thickness of about 110 A. Some chemical analyses were made of isolated membranes after freeing them from salt by precipitating and washing with trichloroacetic acid. Such precipitated membranes consisted predominantly of protein, with little carbohydrate and no peptido-aminopolysaccharide (mucopeptide). Sectioned whole cells of H. halobium contained intracellular electron-opaque structures of unknown function.

scholarly journals Characterization of a calmodulin-dependent high-affinity cyclic AMP and cyclic GMP phosphodiesterase from male mouse germ cells

1984 ◽  
Vol 217 (3) ◽  
pp. 693-700 ◽  
Author(s):  
R Geremia ◽  
P Rossi ◽  
D Mocini ◽  
R Pezzotti ◽  
M Conti
Keyword(s):  
Cyclic Amp ◽  
Germ Cells ◽  
Cyclic Gmp ◽  
Salt Concentration ◽  
Male Mouse ◽  
Competitive Inhibition ◽  
Sedimentation Coefficient ◽  
Phosphodiesterase Activity ◽  
Thermal Stabilities ◽  
High Affinity

Two cyclic nucleotide phosphodiesterase activities were separated by ion-exchange chromatography of cytosol from male mouse germ cells. A form eluted at low salt concentration showed high affinity (Km congruent to 2 microM) and low affinity (Km congruent to 20 microM) for cyclic AMP, and high affinity (Km congruent to 3.5 microM) for cyclic GMP. A second form, eluted at high salt concentration, showed high affinity (Km congruent to 5 microM) for cyclic AMP and was similar to a phosphodiesterase activity described in rat germ cells. The present study was performed to characterize the first form, which represents most of the phosphodiesterase activity in mouse germ cells. The enzyme was sensitive to Ca2+ and calmodulin stimulation, which increased its activity 3-4-fold. Calmodulin stimulation depended on direct interaction of the activator with the enzyme, as indicated by the reversible changes in the chromatographic elution pattern in the presence of Ca2+, as well as by the increase in the sedimentation coefficient in the presence of calmodulin. Reciprocal inhibition kinetics between cyclic AMP and cyclic GMP for the calmodulin-dependent form demonstrated a non-competitive inhibition between the two substrates, suggesting the presence of separate catalytic sites. This is in agreement with kinetic parameters and different thermal stabilities of cyclic AMP- and cyclic GMP-hydrolysing activities. Furthermore, the relevant change in s value, depending on the absence or presence of Ca2+ and calmodulin, suggested that the enzyme is composed of subunits, which aggregate in the presence of the activator. A model for catalytic site composition and reciprocal interaction is also proposed.

scholarly journals Cytosol oestrogen receptor of lactating mammary gland. Effect of heparin on the aggregation of the receptor and interaction of the receptor with heparin-Sepharose

1978 ◽  
Vol 171 (1) ◽  
pp. 137-141 ◽  
Author(s):  
F Auricchio ◽  
A Rotondi ◽  
P Sampaolo ◽  
E Schiavone
Keyword(s):  
Mammary Gland ◽  
Oestrogen Receptor ◽  
High Speed ◽  
Gradient Centrifugation ◽  
Gel Filtration ◽  
Sedimentation Coefficient ◽  
Large Aggregate ◽  
Low Salt ◽  
Lactating Mammary Gland ◽  
Stokes Radius

1. An oestrogen receptor is present in low-salt cytosol of the mammary gland of lactating mice as a large aggregate; it is excluded from gel matrix when filtered on a Sephadex G-200 column and sediments at 7S in sucrose gradients. After incubation of cytosol with heparin, the receptor is dissociated. On a Sephadex G-200 column, it is included in the gel matrix and eluted as a protein with mol.wt. 260000 and a Stokes radius of 6.8nm; it sediments at 6S in sucrose gradients. 2. Dissociation of the mammary-gland cytosol oestrogen receptor seems to be the result of interaction of the oestrogen-receptor complex with heparin. This receptor interacts with heparin covalently bound to Sepharose, thereafter sedimenting at 6S. By using this interaction, the cytosol receptor was purified 200-fold compared with the homogenate, with a yield of 70%. 3. The cytosol receptor that was not incubated or was incubated with heparin was much smaller during sucrose-gradient centrifugation than during gel filtration. This discrepancy can be explained by pressure-induced dissociation during high-speed centrifugation. This possibility is supported by the decrease in the sedimentation coefficient of the receptor with increased duration of centrifugation.

Maintenance of Photophosphorylation Despite Inhibition of the Transthylakoid pH Gradient by Tetracaine

1992 ◽  
Vol 47 (9-10) ◽  
pp. 717-725 ◽  
Author(s):  
Henrik Laasch
Keyword(s):  
Salt Concentration ◽  
Spinacia Oleracea ◽  
Electron Flow ◽  
Energy Coupling ◽  
Ph Gradient ◽  
Proton Pumps ◽  
Linear Electron ◽  
Ps Ii ◽  
Spinacia Oleracea L ◽  
Low Salt

The inhibition of the transthylakoid pH gradient, ΔpH, and of photophosphorylation by the local anesthetic tetracaine was investigated with isolated chloroplasts from Spinacia oleracea L. Tetracaine strongly inhibited ΔpH in the presence of low salt concentrations. In the presence of high salt concentrations, the inhibition of ΔpH was much smaller. This effect of salt concentration was observed only when both, cation and anion were easily membrane permeable. It was concluded that the effect of salts on ΔpH inhibition was excerted on the inside of the thylakoid membrane. The rate of photophosphorylation, Vp, driven by the PS Idependent artificial proton carrier phenazine methosulfate decreased with ΔpH in the presence of both, high and low salt concentrations. In contrast, Vp driven by the endogenous proton pumps of PS II + I-dependent linear electron flow was largely independent of ΔpH changes in the presence of low salt concentration. It appeared that energy coupling during linear electron transport, in contrast to artificially produced PS I-dependent coupling, may be localized to membrane-bound proton domains which are not accessible to the employed indicators of ΔpH. The data were discussed with respect to recent hypotheses on localized energy coupling in chloroplasts.

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