scholarly journals Protein diffusion through charged nanopores with different radii at low ionic strength

2014 ◽  
Vol 16 (39) ◽  
pp. 21570-21576 ◽  
Author(s):  
Pieter Stroeve ◽  
Masoud Rahman ◽  
Lekkala Dev Naidu ◽  
Gilbert Chu ◽  
Morteza Mahmoudi ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
Protein Diffusion ◽  
Similar Molecular Weight ◽  
Low Ionic Strength ◽  
Charged Membranes ◽  
Pore Permeability

Pore permeability for two similar molecular weight proteins (BSA and BHb) through nanoporous charged membranes at low ionic strength (I = 0.001 M).

Human and Bovine Plasminogen-Free Thrombin, Purified by Means of Gel Filtration and Ion Exchange Chromatography

1966 ◽  
Vol 15 (03/04) ◽  
pp. 501-510 ◽  
Author(s):  
W Berg ◽  
K Korsan-Bengtsen ◽  
J Ygge
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
Large Scale ◽  
Gel Filtration ◽  
Trisodium Citrate ◽  
Exchange Chromatography ◽  
Simple Method ◽  
Commercial Preparation ◽  
Usual Manner ◽  
Low Ionic Strength

SummaryA simple method for preparation of plasminogen-free human and bovine thrombin is described.Crude thrombin was prepared in the usual manner from oxalated plasma by means of adsorption on BaSO4, elution with trisodium citrate and activating the eluate from BaSO4 with tissue thromboplastin.This crude thrombin was purified by means of gel-filtration and chromatography on CM-Sephadex A-50.The gel-filtration was performed on three types of Sephadex, G-75, G-50, and G--25. By means of Sephadex G-75 the thrombin was well separated from the main part of inert protein and this type of Sephadex was used for the purification in large-scale. Separation of thrombin from protein of higher molecular weight was also obtained with Sephadex G-50 but not with Sephadex G-25 indicating a molecular weight of thrombin between 4000 and 10,000.The importance of using an elution buffer of sufficient high ionic strength for gel-filtration is shown. A great deal of the thrombin was adsorbed to the Sephadex if the gel-filtration was performed at a too low ionic strength.The final preparation contained 30,000 NIH units of thrombin per mg tyrosin and no detectable plasminogen.The commercial preparation “Topostasine” was also purified in the same manner, but the plasminogen content in “Topostasine” was high and could not be completely separated from thrombin.

scholarly journals Production and characterisation of a marine Halomonas surface-active exopolymer

2019 ◽  
Vol 104 (3) ◽  
pp. 1063-1076
Author(s):  
Tony Gutierrez ◽  
Gordon Morris ◽  
Dave Ellis ◽  
Barbara Mulloy ◽  
Michael D. Aitken
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
High Molecular Weight ◽  
Positive Influence ◽  
Contaminated Sites ◽  
Crude Oils ◽  
Magnetic Resonance Spectroscopic Analysis ◽  
Surface Active ◽  
Low Ionic Strength ◽  
Pure Hydrocarbon

AbstractDuring screening for novel emulsifiers and surfactants, a marine gammaproteobacterium, Halomonas sp. MCTG39a, was isolated and selected for its production of an extracellular emulsifying agent, P39a. This polymer was produced by the new isolate during growth in a modified Zobell’s 2216 medium amended with 1% glucose, and was extractable by cold ethanol precipitation. Chemical, chromatographic and nuclear magnetic resonance spectroscopic analysis confirmed P39a to be a high-molecular-weight (~ 261,000 g/mol) glycoprotein composed of carbohydrate (17.2%) and protein (36.4%). The polymer exhibited high emulsifying activities against a range of oil substrates that included straight-chain aliphatics, mono- and alkyl- aromatics and cycloparaffins. In general, higher emulsification values were measured under low (0.1 M PBS) compared to high (synthetic seawater) ionic strength conditions, indicating that low ionic strength is more favourable for emulsification by the P39a polymer. However, as observed with other bacterial emulsifying agents, the polymer emulsified some aromatic hydrocarbon species, as well as refined and crude oils, more effectively under high ionic strength conditions, which we posit could be due to steric adsorption to these substrates as may be conferred by the protein fraction of the polymer. Furthermore, the polymer effected a positive influence on the degradation of phenanthrene by other marine bacteria, such as the specialist PAH-degrader Polycyclovorans algicola. Collectively, based on the ability of this Halomonas high-molecular-weight glycoprotein to emulsify a range of pure hydrocarbon species, as well as refined and crude oils, it shows promise for the bioremediation of contaminated sites.

Properties and characteristics of a bacteriocin-like substance produced by Propionibacterium acnes isolated from dental plaque

1988 ◽  
Vol 34 (12) ◽  
pp. 1344-1347 ◽  
Author(s):  
Greg E. Paul ◽  
S. James Booth
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
Dental Plaque ◽  
Propionibacterium Acnes ◽  
Gram Positive ◽  
Gram Negative ◽  
A Cell ◽  
Approximate Molecular Weight ◽  
Low Ionic Strength

A cell-associated bacteriocinlike substance with an approximate molecular weight of 78 000 was isolated from an oral isolate of Propionibacterium acnes. The substance was bacteriostatic and was active against both gram-positive and gram-negative anaerobes. Lysozyme inhibited the activity of the bacteriocinlike substance at low ionic strength.

scholarly journals The behaviour of heterogeneous nuclear ribonucleic acid in partially and completely denaturing conditions

1974 ◽  
Vol 141 (2) ◽  
pp. 477-484 ◽  
Author(s):  
Michael E. Bramwell
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
High Temperatures ◽  
Sedimentation Rate ◽  
Ribonucleic Acid ◽  
28S Rrna ◽  
Nuclear Rna ◽  
Step Down ◽  
Low Ionic Strength ◽  
Nucleolar Rna

It is shown that the heterogeneous nuclear RNA (HnRNA) synthesized in the presence of actinomycin and at low and high temperatures sediments in low-ionic-strength sucrose gradients between the rRNA components, similar to the unmethylated RNA synthesized under ‘step-down’ conditions. If the ionic strength is increased then the HnRNA sediments more rapidly than 28S rRNA, with a large proportion about the 45S precursor rRNA position. Initially this was thought to be due to aggregation of the HnRNA; however, centrifugation and electrophoresis in completely denaturing conditions suggest that the molecular weight of this species of RNA is very large The experiments reveal that HnRNA is conformationally unstable relative to the nucleolar RNA and that the slower sedimentation rate of HnRNA in 5mm-EDTA–Tris base–sucrose gradients reflects the greater expansion of the HnRNA relative to the nucleolar RNA. The implications of this finding are discussed.

Subunit Structure of Human Factor VIII

1975 ◽  
Author(s):  
J. A. van Mourik ◽  
W. T. LaBruyère ◽  
I. A. Mochtar
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
Factor Viii ◽  
Apparent Molecular Weight ◽  
Subunit Structure ◽  
Single Chain ◽  
Covalent Bonds ◽  
Single Polypeptide Chain ◽  
Human Factor Viii ◽  
Low Ionic Strength

Several investigations have shown that factor VIII is a high molecular weight aggregate and that both non-covalent and disulphide bonds contribute to the stabilization of the macromolecular factor VIII aggregate. As shown previously (1) disruption of non-covalent bonds can be accomplished at relatively low ionic strength and neutral pH and results via a series of homologous oligomers (having constant charge/mass ratios) in 2 immunologically non-related subunits. Thus, the generally accepted concept that factor VIII is constructed of identical subunits seems incorrect. For several reasons we assume that the observed fragmentation at low ionic strength is not due to proteolytic breakdown. However, this view is not favoured by the observation that tryptic and plasmic digestion of factor VIII (in aggregated form) results in gel electrophoresis patterns comparable with those obtained of factor VIII after low ionic strength dissociation. Further, evidence will be presented that the assumption that reduction of factor VIII under denaturating conditions results in a single polypeptide chain is also no longer tenable. As far as the reduction of aggregated factor VIII is concerned our observations agree with data from the literature, that is, reduction of factor VIII results in 1 major subunit with an apparent molecular weight of approximately 270.000. However, when factor VIII is first dissociated at low ionic strength followed by reduction, smaller fragments appear in the electrophoresis pattern. Thus, it seems most likely that the apparent single chain subunit is in fact, constructed of smaller non-covalently linked fragments.(1) J. A. van Mourik et al. Thrombosis Research, 4, 155, 1974.

scholarly journals Chromatography of plasma proteins on immobilized Cibacron Blue F3-GA. Mechanism of the molecular interaction

1982 ◽  
Vol 203 (3) ◽  
pp. 637-641 ◽  
Author(s):  
E Gianazza ◽  
P Arnaud
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
Molecular Interaction ◽  
Plasma Proteins ◽  
Hydrophobic Interactions ◽  
Molecular Characteristics ◽  
Acidic Ph ◽  
Cibacron Blue ◽  
Ligand Affinity ◽  
Low Ionic Strength

Fractionation of plasma proteins on immobilized Cibacron Blue F3-GA (Affi-gel Blue) under different conditions of pH, ionic strength and temperature was studied. At acidic pH the unbound proteins were eluted in order of increasing pI (the Affi-gel Blue behaving as ion-exchanger); at basic pH and at low ionic strength they were eluted in order of decreasing molecular weight (separation by diffusion-exclusion). For the proteins that were either retarded in comparison with substances of similar molecular characteristics, or that were bound to the resin, pseudo-ligand affinity or hydrophobic interactions were also implicated.

scholarly journals Dictyostelium myosin: characterization of chymotryptic fragments and localization of the heavy-chain phosphorylation site.

1981 ◽  
Vol 89 (1) ◽  
pp. 104-108 ◽  
Author(s):  
G Peltz ◽  
E R Kuczmarski ◽  
J A Spudich
Keyword(s):  
Molecular Weight ◽  
Ionic Strength ◽  
Atpase Activity ◽  
Heavy Chain ◽  
Polyacrylamide Gel Electrophoresis ◽  
Phosphorylation Site ◽  
Polyacrylamide Gel ◽  
Maximal Velocity ◽  
Dictyostelium Myosin ◽  
Low Ionic Strength

Chymotrypsin cleaves Dictyostelium myosin in half, splitting the heavy chain (210,000 daltons) into two fragments of 105,000 daltons each. One of the two major fragments is soluble at low ionic strength and has a native molecular weight of 130,000. As judged by SDS polyacrylamide gel electrophoresis, this soluble fragment consists of the two intact myosin light chains of 18,000 and 16,000 daltons and a 105,000-dalton polypeptide derived from the myosin heavy chain. The soluble fragment retains actin-activated ATPase activity and the ability to bind to actin in an ATP-dissociable fashion. The maximal velocity of the actin-activated ATPase activity of the soluble fragment is 80% of that of uncleaved myosin, although its apparent Km for actin is 12-fold greater than that of myosin. In addition to the major soluble 105,000-dalton fragment discussed above, chymotryptic cleavage of the Dictyostelium myosin also generates fragments that are insoluble at low ionic strength. The major insoluble fragment is 105,000 daltons on an SDS polyacrylamide gel and forms thick filaments that are devoid of myosin heads. A less prevalent insoluble fragment has a molecular weight of 83,000 and is probably a subfragment of the insoluble 105,000-dalton fragment. The heavy chain of myosin is phosphorylated in vivo and the phosphorylation site has been localized to the insoluble fragments, which derive from the tail portion of the myosin molecule.

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