Rat α2 acute-phase macroglobulin. Isolation and physicochemical properties

1. Rat α2 acute-phase macroglobulin was isolated from turpentine-injected rats by Sephadex G-200 chromatography and ion-exchange chromatography on DEAE-cellulose. This method, since it does not include (NH4)2SO4 treatment, allows the study of the physicochemical as well as the biological properties of the molecule. 2. The purity of the preparation was demonstrated by ultracentrifugation, polyacrylamide-gel electrophoresis, fused “rocket” immunoelectrophoresis as well as double immunodiffusion. 3. The rat α2 acute-phase macroglobulin was characterized in terms of its main physical and chemical properties. Its isoelctric point was determined by isoelectrofocusing to be 4.55; s020,w was 18.4S and E1%/1cm at 278 nm was 6.8. The mol.wt. was determined by light-scattering to be 770000. 4. The amino acid content was compared with that of rat α1 macroglobulin and was found very similar. The carbohydrate composition of α2 acute-phase macroglobulin was determined to be: hexose, 4.25%; glucosamine, 3.4%; sialic acid, 2%; fucose, 0.2%. From these results it was concluded that α2 acute-phase macroglobulin, although a typical acute-phase reactant, possesses the characteristic physicochemical properties of α macroglobulins.

Download Full-text

Purification of a Highly Potent Heparin-Like Anticoagulant from Healthy Human Plasma

10.1055/s-0039-1684469 ◽

1979 ◽

Author(s):

Robert H. Yue ◽

Toby Starr ◽

Menard M. Gertler

Keyword(s):

Human Plasma ◽

Specific Activity ◽

Anticoagulant Activity ◽

Polyacrylamide Gel Electrophoresis ◽

Deae Cellulose ◽

Chemical Properties ◽

Healthy Human ◽

Naturally Occurring ◽

Salt Gradient ◽

Physical And Chemical

A highly potent-heparin-like anticoagulant (accelerator) has been purified from citrated healthy human plasma. After heat defibrination and BaSO4. treatment on the plasma, the accelerator was adsorbed onto a DEAE-cellulose column and elution was achieved using a high ionic strength linear buffered salt gradient. The eluted accelerator was further purified by polyacrylamide gel electrophoresis. The purified accelerator can accelerate the inhibition of thrombin by antithrombin III and has a specific activity of 226 heparin units/mg of carbohydrate. The accelerator contains a small amount of protein-like material. The amount of accelerator present in healthy human blood is extremely small and can only be first detected in the concentrate after the DEAE-cellu-lose chromatography. A mere 0.5 mg of the purified accelerator is obtained from 100 ml of human plasma. Concomitant with this investigation, a second heparin-like substance also has been purified but has very low anticoagulant activity in terms of heparin units. The naturally occurring accelerator may function as heparin in the circulating blood and its level in blood may have a clinical significance in thrombotic vascular disease. Further work on its physical and chemical properties is now in progress.

Download Full-text

Physicochemical characterisation of kafirins extracted from sorghum grain and dried distillers grain with solubles related to their biomaterial functionality

Scientific Reports ◽

10.1038/s41598-021-94718-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Umar Shah ◽

Deepak Dwivedi ◽

Mark Hackett ◽

Hani Al-Salami ◽

Ranjeet P. Utikar ◽

...

Keyword(s):

Secondary Structure ◽

Physicochemical Properties ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Chemical Properties ◽

X Ray ◽

Distillers Grain ◽

Sorghum Grain ◽

Dried Distillers Grain ◽

Polypeptide Profile

AbstractKafirin, the hydrophobic prolamin storage protein in sorghum grain is enriched when the grain is used for bioethanol production to give dried distillers grain with solubles (DGGS) as a by-product. There is great interest in DDGS kafirin as a new source for biomaterials. There is however a lack of fundamental understanding of how the physicochemical properties of DDGS kafirin having been exposed to the high temperature conditions during ethanol production, compare to kafirin made directly from the grain. An understanding of these properties is required to catalyse the utilisation of DDGS kafirin for biomaterial applications. The aim of this study was to extract kafirin directly from sorghum grain and from DDGS derived from the same grain and, then perform a comparative investigation of the physicochemical properties of these kafirins in terms of: polypeptide profile by sodium-dodecyl sulphate polyacrylamide gel electrophoresis; secondary structure by Fourier transform infra-red spectroscopy and x-ray diffraction, self-assembly behaviour by small-angle x-ray scattering, surface morphology by scanning electron microscopy and surface chemical properties by energy dispersive x-ray spectroscopy. DDGS kafirin was found to have very similar polypeptide profile as grain kafirin but contained altered secondary structure with increased levels of β-sheets. The structure morphology showed surface fractals and surface elemental composition suggesting enhanced reactivity with possibility to endow interfacial wettability. These properties of DDGS kafirin may provide it with unique functionality and thus open up opportunities for it to be used as a novel food grade biomaterial.

Download Full-text

Structure Determination of C23H19N4OBr from Synchrotron Data

Acta Crystallographica Section A Foundations and Advances ◽

10.1107/s2053273314098581 ◽

2014 ◽

Vol 70 (a1) ◽

pp. C141-C141

Author(s):

Ozen Ozgen ◽

Engin Kendi ◽

Semra Koyunoglu ◽

Akgul Yesilada ◽

Hwo-Shuenn Sheu

Keyword(s):

Powder Diffraction ◽

Structural Information ◽

Chemical Properties ◽

Biological Properties ◽

Cell Parameters ◽

Fundamental Understanding ◽

Physical And Chemical ◽

Radiation Research ◽

And Chemical Properties

A significant part of medicine is based on the discovery and development of drugs. It is very important to know the crystal structure of pharmaceutical compounds for fundamental understanding of structure, physical and chemical properties. Many of these materials are available only as powders. So any structural information must be obtained from powder diffraction. I am going to present following the stages while solving the structure of C23H19N4OBr, 2-[3-phenyl-4(m-bromophenyl)-2-pyrazolin-1-yl]-3-methyl-4(3H)-quinazolinone, from 2-pyrazolines derivatives. The compounds are known to display various biological properties such as fungicidal insecticidal, anti bacterial, anti viral activities, pharmacological properties such as antiinflammatory agents and have industral properties(1). The powder diffraction data was collected with Debye Scherrer camera at the BL01C2 beamline at room temperature in National Synchrotron Radiation Research Center(NSRRC), Taiwan. X-ray of wavelength was 1.0333Å. This compound crystallizes in orthorhombic system space group P bca, Z=8, unit cell parameters of a=25.83(1)Å, b=15.55(5)Å, c=10.63(3)Å, and V=4266.0(10)Å3. Reliability factors were reached Rwp=0.075, Rp=0.053, RB=0.086 ve S=1.31 after Rietveld refinement.

Download Full-text

Comparing the Physicochemical Properties of Upgraded Biomass Fuel by Torrefaction and the Ashless Technique

Applied Sciences ◽

10.3390/app9245519 ◽

2019 ◽

Vol 9 (24) ◽

pp. 5519 ◽

Cited By ~ 2

Author(s):

Lkhagvadorj Sh ◽

Byoung-Hwa Lee ◽

Young-Joo Lee ◽

Chung-Hwan Jeon

Keyword(s):

Physicochemical Properties ◽

Chemical Properties ◽

High Heating Rate ◽

Pine Sawdust ◽

Herbaceous Biomass ◽

High Heating ◽

Pitch Pine ◽

Proximate And Ultimate Analysis ◽

Physical And Chemical ◽

And Chemical Properties

The purpose of this study was to investigate and compare the influence of torrefaction and an ashless process on the physical and chemical properties of pitch pine sawdust (PSD) and kenaf as types of woody and herbaceous biomass. The physicochemical properties of the materials pretreated by the ashless process with torrefaction including proximate and ultimate analysis, hydrophobicity, grindability, morphology, and structure were analyzed. The results showed that when ashless Kenaf was torrefied, the high heating rate and atomic ratios of O/C and H/C increased. The tendency of the torrefied, ashless Kenaf to absorb water decreased, and it became more hydrophobic (approximately 0% for the uptake rate of moisture). In addition, the grindability of the torrefied, ashless Kenaf was substantially improved compared to that of pretreated PSD. Brunauer–Emmett–Teller and scanning electron microscopy results showed that when Kenaf was pretreated, particles easily lost their fibrous structure and cracked as the number of macropores decreased. These results indicate that the herbaceous biomass of Kenaf, when pretreated with both torrefaction and the ashless process, exhibits improved physicochemical properties compared to the woody PSD.

Download Full-text

Comparison of earthworm populations in arable and grassland fields in the Outer Western Carpathians, South Poland

Biologia ◽

10.1515/biolog-2016-0035 ◽

2016 ◽

Vol 71 (3) ◽

Cited By ~ 4

Author(s):

Agnieszka Józefowska ◽

Anna Miechówka ◽

Jan Frouz

Keyword(s):

Chemical Properties ◽

Biological Properties ◽

Outer Western Carpathians ◽

Geographical Regions ◽

Type Of Land Use ◽

Earthworm Biomass ◽

The Mean ◽

Physical And Chemical ◽

The Impact ◽

Earthworm Populations

AbstractThe impact of different geographical regions (Silesian Foothills, region 1 and Maly Beskids, region 2), and method of soil use (arable field and grassland) on the main soil properties and biological activity was studied. Earthworm biomass, density and diversity, as well as dehydrogenase activity, were analysed. Significant soil physical and chemical properties were more affected by regions, whereas the type of land use had a greater impact on the biological properties. The mean earthworm density was 213 ind. m

Download Full-text

Mouse Uterine Alkaline Phosphatase: Improved Purification by Affinity Chromatography and Further Characterization of the Enzyme

Australian Journal of Biological Sciences ◽

10.1071/bi9800279 ◽

1980 ◽

Vol 33 (3) ◽

pp. 279 ◽

Cited By ~ 3

Author(s):

RN Murdoch ◽

Louise E Buxton ◽

DJ Kay

Keyword(s):

Alkaline Phosphatase ◽

Affinity Chromatography ◽

Polyacrylamide Gel Electrophoresis ◽

Gel Filtration ◽

Deae Cellulose ◽

Apparent Molecular Weight ◽

Anion Exchange Chromatography ◽

Pregnant Mouse ◽

Exchange Chromatography ◽

Enzyme Preparations

An improved procedure for the purification of alkaline phosphatase from about 10 g of day 7 pregnant mouse uterine tissue is described. Following homogenization, the procedure involved solubilization and extraction with 0�8% (v/v) Triton X-lOO and 20% (v/v) n-butanol, ammonium sulfate precipitation, concanavalin A-Sepharose 4B affinity chromatography, DEAE-cellulose anion-exchange chromatography and Sephacryl S200 gel filtration. On subjecting 2162-fold purified enzyme preparations to polyacrylamide-gel electrophoresis, a single band of protein coincident with the zone of enzyme activity and having an apparent molecular weight of 205 OOO� lOOOO was identified. Affinity chromatography yielded the largest increase in purity of any step in the procedure and established the glycoprotein nature of the uterine enzyme.

Download Full-text

PURIFICATION OF "INHIBIN" FROM HUMAN OVARIAN FOLLICULAR FLUID

Acta Endocrinologica ◽

10.1530/acta.0.0900157 ◽

1979 ◽

Vol 90 (1) ◽

pp. 157-166 ◽

Cited By ~ 48

Author(s):

S. Chari ◽

C. R. N. Hopkinson ◽

E. Daume ◽

G. Sturm

Keyword(s):

Follicular Fluid ◽

Polyacrylamide Gel Electrophoresis ◽

Gel Filtration ◽

Deae Cellulose ◽

Exchange Chromatography ◽

Male Rats ◽

Acetone Powder ◽

Gel Chromatography ◽

Apparent Homogeneity ◽

Ovarian Follicular Fluid

ABSTRACT Following the earlier demonstration of inhibin-like activity in human ovarian follicular fluid a method for its purification to apparent homogeneity is described. The fluid was converted to acetone powder and subjected sequentially to ammonium sulphate fractionation, gel chromatography on Sephadex G-200, continuous gradient ion-exchange chromatography on DEAE-cellulose, first with a pH gradient from 8.0 to 4.0 and then with a NaCl gradient to 1 m at pH 5.2. The active fraction from this step was subjected to gel filtration on Sephadex G-100 and finally passed through an Amicon Centriflo membrane CF-25 (cut off point: 25 000 m.w.). The ultrafiltrate was homogeneous by SDS-polyacrylamide gel electrophoresis, had a molecular weight of the order of 23 000 and was capable of suppressing serum gonadotrophin levels in the castrated male rats in as low a dose as 25 μg/rat.

Download Full-text

Isolation and partial characterization of canine epidermal growth factor/urogastrone

Biochemical Journal ◽

10.1042/bj2290611 ◽

1985 ◽

Vol 229 (3) ◽

pp. 611-619 ◽

Cited By ~ 8

Author(s):

R Kobayashi ◽

J R Reeve ◽

J H Walsh

Keyword(s):

Growth Factor ◽

Epidermal Growth Factor ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Deae Cellulose ◽

Exchange Chromatography ◽

Mouse Fibroblast ◽

A431 Cells ◽

Peptide Component ◽

Epidermal Growth

Canine epidermal growth factor (EGF)/urogastrone was partially purified from dog urine by fractional precipitation with (NH4)2SO4, ion-exchange chromatography with DEAE-cellulose DE-52, gel filtration with Sephadex G-50, and a second DE-52 chromatography, to yield receptor-competing activity equivalent to 13 micrograms of standard mouse EGF/litre of starting urine. The purification was monitored by a competitive radioreceptor assay using fixed monolayers of A431 cells. The partially purified canine EGF/urogastrone demonstrated a growth-stimulating activity in 3T3 mouse fibroblast cells as potent as mouse EGF. Analysis by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis revealed one major peptide component with an Mr similar to that of mouse EGF, and two minor peptides of slightly higher Mr. The major peptide component was isolated after reduction and its amino acid composition was determined.

Download Full-text

Purification and characterization of the extracellular α-amylase activity of the yeast Schwanniomyces alluvius

Biochemistry and Cell Biology ◽

10.1139/o87-116 ◽

1987 ◽

Vol 65 (10) ◽

pp. 899-908 ◽

Cited By ~ 21

Author(s):

F. Moranelli ◽

M. Yaguchi ◽

G. B. Calleja ◽

A. Nasim

Keyword(s):

Amino Acid ◽

Amylase Activity ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Gel Filtration ◽

Deae Cellulose ◽

Exchange Chromatography ◽

Proteinase K ◽

Ph Optimum ◽

Sds Page

The extracellular α-amylase activity of the yeast Schwanniomyces alluvius has been purified by anion-exchange chromatography on DEAE-cellulose and gel-filtration chromatography on Sephadex G-100. Sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS–PAGE) and N-terminal amino acid analysis of the purified sample indicated that the enzyme preparation was homogeneous. The enzyme is a glycoprotein having a molecular mass of 52 kilodaltons (kDa) estimated by SDS–PAGE and 39 kDa by gel filtration on Sephadex G-100. Chromatofocusing shows that it is an acidic protein. It is resistant to trypsin but sensitive to proteinase K. Its activity is inhibited by the divalent cation chelators EDTA and EGTA and it is insensitive to sulfhydryl-blocking agents. Exogenous divalent cations are inhibitory as are high concentrations of monovalent salts. The enzyme has a pH optimum between 3.75 and 5.5 and displays maximum stability in the pH range of 4.0–7.0. Under the conditions tested, the activity is maximal between 45 and 50 °C and is very thermolabile. Analysis of its amino acid composition supports its acidic nature.

Download Full-text

Nanostructured terbium-doped ceria spheres: effect of dopants on their physical and chemical properties under reducing and oxidizing conditions

Journal of Materials Chemistry A ◽

10.1039/c5ta02535d ◽

2015 ◽

Vol 3 (31) ◽

pp. 16120-16131 ◽

Cited By ~ 8

Author(s):

L. M. Acuña ◽

F. F. Muñoz ◽

C. A. Albornoz ◽

A. G. Leyva ◽

R. T. Baker ◽

...

Keyword(s):

Physicochemical Properties ◽

Synthesis Method ◽

Chemical Properties ◽

Physical And Chemical Properties ◽

Doped Ceria ◽

Physical And Chemical ◽

And Chemical Properties

The effect of Tb content and synthesis method on the physicochemical properties of nanostructured Tb-doped ceria spheres was studied. The nanostructured spheres contained more Tb as Tb3+ than conventionally prepared nanopowders.

Download Full-text