Coated vesicles from chicken liver bind ferritin

1989 ◽  
Vol 92 (2) ◽  
pp. 187-196
Author(s):  
A. Passaniti ◽  
T.F. Roth
Keyword(s):  
Glass Bead ◽  
Coated Vesicles ◽  
Chicken Liver ◽  
Iron Storage ◽  
Controlled Pore Glass ◽  
Pore Glass ◽  
Detergent Treatment ◽  
Liver Ferritin ◽  
Triton X ◽  
Iron Storage Protein

The iron-storage protein ferritin was found to be associated with highly purified coated vesicles (CV) from chicken liver. Chicken liver ferritin was morphologically similar to ferritin from horse spleen and could be isolated using a specific anti-ferritin monoclonal antibody. This antibody recognized a 240 X 10(3) Mr form of chicken ferritin but not the 22 X 10(3) Mr ferritin subunit after protein transfer to nitrocellulose. CV purified by controlled-pore glass-bead chromatography also contained ferritin when assayed by monoclonal anti-ferritin antibody using a sensitive enzyme-linked assay. Ferritin remained associated with CV even after re-chromatography. Ferritin particles were observed to be associated with CV by electron microscopy. CV-associated ferritin could be quantitatively removed from CV by treatment of the CV with 0.5 M-Tris-HC1 + 2M-urea at pH 8.5, conditions that also lead to dissociation of the clathrin lattice. Triton X-100 detergent treatment did not affect the association of ferritin with CV. These results indicate that purified CV from chicken liver contain ferritin in association with the clathrin lattice. The possible functional significance of this association is discussed.

scholarly journals Purification of chicken liver ferritin by two novel methods and structural comparison with horse spleen ferritin

1989 ◽  
Vol 258 (2) ◽  
pp. 413-419 ◽  
Author(s):  
A Passaniti ◽  
T F Roth
Keyword(s):  
Gel Filtration ◽  
Average Diameter ◽  
Chicken Liver ◽  
Two Dimensional Gel Electrophoresis ◽  
Controlled Pore Glass ◽  
Isoelectric Points ◽  
Pore Glass ◽  
The Difference ◽  
Liver Ferritin ◽  
Horse Spleen Ferritin

Ferritin was purified from chicken liver by two different methods: gel filtration on controlled-pore glass beads, and immunoaffinity chromatography employing a chicken ferritin-specific monoclonal antibody that did not cross-react with horse spleen ferritin. This antibody recognizes intact ferritin and an oligomeric 240 kDa form of the molecule after protein transfer to nitrocellulose, but not the 22 kDa chicken ferritin subunit. Chicken liver ferritin purified by these methods exhibited reduced migration on non-denaturing polyacrylamide gels compared with horse spleen ferritin. These results were consistent with the difference in calculated isoelectric points of chicken and horse ferritin subunits. By two-dimensional gel electrophoresis, chicken ferritin 22 kDa subunits exhibited isoelectric points from 6.1 to 6.6 whereas horse spleen ferritin subunits exhibited isoelectric points of 5.8-6.3. The 240 kDa form of the chicken ferritin molecule had an isoelectric point of 6.6 whereas the 210 kDa form of the horse ferritin molecule had isoelectric points of 5.1 and 4.9. Intact chicken liver ferritin particles were 13.4 +/- 0.8 nm (controlled-pore glass-purified) and 12.5 +/- 0.9 nm (affinity-purified) in diameter when viewed by electron microscopy. Horse spleen ferritin consisted of slightly smaller particles with an average diameter of 11.0 +/- 0.7 nm. However, ferritin from chicken liver and horse spleen co-migrated with an apparent molecular mass of 470 kDa when analysed by Sepharose 4B gel filtration chromatography. These results indicate that, consistent with results from other published purification methods, the chicken ferritin purified by the methods reported here exhibits both structural similarities to, and differences from, horse spleen ferritin.

scholarly journals Tubulin as a molecular component of coated vesicles.

1983 ◽  
Vol 97 (4) ◽  
pp. 1191-1199 ◽  
Author(s):  
W G Kelly ◽  
A Passaniti ◽  
J W Woods ◽  
J L Daiss ◽  
T F Roth
Keyword(s):  
Microscopic Analysis ◽  
Bovine Brain ◽  
Coated Vesicles ◽  
Antigenic Determinants ◽  
Molecular Component ◽  
Electron Microscopic ◽  
Molecular Weights ◽  
Controlled Pore Glass ◽  
Sds Page ◽  
Pore Glass

Two proteins of 53,000 and 56,000 mol wt have been found to be associated with coated vesicles (CV) purified from bovine brain and chicken liver. These proteins share molecular weights, isoelectric points, and antigenic determinants with alpha- and beta-tubulins purified from bovine brain. Based on SDS PAGE and electron microscopic analysis of controlled pore glass bead exclusion column fractions, both the tubulins and the major CV polypeptide clathrin were found to chromatograph as components of a single kinetic particle. In addition, tubulin and CV antigens assayed by a sensitive enzyme-linked-immunoadsorbent method eluted from the columns with constant stoichiometry. These data provide evidence that tubulin is a molecular component of coated vesicles.

A controlled pore glass bead assay for the measurement of cytoplasmic and nuclear glucocorticoid receptors

1985 ◽  
Vol 22 (6) ◽  
pp. 693-698 ◽  
Author(s):  
Glenn J. Gormley ◽  
Verne D. Hospelhorn ◽  
Mohamed K. Khan ◽  
Elwood V. Jensen
Keyword(s):  
Glass Bead ◽  
Glucocorticoid Receptors ◽  
Controlled Pore Glass ◽  
Pore Glass

Polygonal profiles of ferritin molecules

1983 ◽  
Vol 41 ◽  
pp. 600-601
Author(s):  
William H. Massover
Keyword(s):  
Axial Ratio ◽  
X Ray Diffraction ◽  
Rhombic Dodecahedron ◽  
Apparent Contradiction ◽  
Iron Storage ◽  
X Ray ◽  
Hexagonal Shape ◽  
Axes Of Symmetry ◽  
Apparent Conflict ◽  
Iron Storage Protein

The molecular structure of the iron-storage protein, ferritin, is becoming known in ever finer detail. The 24 apoferritin subunits (MW ca. 20,000) have a 2:1 axial ratio and are polymerized with 4:3:2 symmetry to form an outer shell surrounding a variable amount of microcrystalline iron, Recent x-ray diffraction results indicate that the projected outline of the native molecule has a quasi-hexagonal shape when viewed down the 3-fold axes of symmetry, and a quasi-square shape when looking down the 4-fold axes. To date, no electron microscope study has reported observing anything other than circular profiles, which would indicate that ferritin is strictly spherical. The apparent conflict between the "hollow sphere" of electron microscopy (E.M.) and the "truncated rhombic dodecahedron" of x-ray diffraction could reflect the poorer effective resolution of E.M. coming from radiation damage, staining, drying, etc. The present study investigates the detailed shape of individual ferritin molecules in order to search for the predicted aspherical profiles and to interpret the nature of this apparent contradiction.

Calorimetric Sensor for Ethanol Using Ni2+-nitrilotriacetic Acid (NTA) Resin Immobilized Alcohol Dehydrogenase (ADH)

2020 ◽  
Vol 16 (6) ◽  
pp. 795-799
Author(s):  
YongJin Li
Keyword(s):  
Cost Effective ◽  
Nitrilotriacetic Acid ◽  
Repeated Measurements ◽  
Great Promise ◽  
Calorimetric Sensor ◽  
Controlled Pore Glass ◽  
Pore Glass ◽  
One Step ◽  
Immobilization Matrix ◽  
Enzyme Thermistor

Background: A simple, fast and economic analytical method for the determination of ethanol is important for clinical, biological, forensic and physico-legal purposes. Methods: Ni2+-NTA resin was used as an immobilization matrix for the simple one-step purification/ immobilization of his6-tagged ADH. Different alcohols with a concentration range of 0.5-50% V/V, namely methanol, ethanol and propanol were measured using prepared ADH enzyme thermistor. The ethanol content of Tsingtao beer was tested as a real sample containing alcohol. Reproducibility and stability of prepared ADH enzyme thermistor were also investigated by repeated measurements. Results: In comparison to the controlled pore glass (a common used support for the immobilization of enzyme) used in thermal biosensor, the use of Ni2+-NTA resin not only led to simple one-step purification/ immobilization by his6-tagged ADH binding to Ni2+-NTA resin, but also made the immobilizing supports reusable. The prepared biosensor can be used to determine ethanol and methanol by the calorimetric measurement. A linear range of 1 -32% (V/V) and 2-20% (V/V) was observed for ethanol and methanol, respectively. The detection limits were 0.3% (V/V) and 1% (V/V) for ethanol and methanol, respectively. The tested ethanol concentration of Tsingtao beer was 4.5% V/V, which is comparable with the labeled alcohol by volume (ABV) 4.80%. Conclusion: Ni2+-NTA resin, as an immobilization matrix in ET sensor, provides a simple one-step purification/immobilization for His6-tagged recombinase and a reusable immobilization matrix. The prepared biosensor exhibits good repeatability and stability. Such a new biosensor shows great promise for rapid, simple, and cost-effective analysis of ethanol and methanol, both in qualitative and in quantitative tests.

Improved Conditions for Solid Phase Synthesis of Oligonucleotides on PS-PEG Copolymers

1995 ◽  
Vol 50 (7) ◽  
pp. 1096-1100 ◽  
Author(s):  
Ernst Bayer ◽  
Konrad Bleicher ◽  
Martin Maier
Keyword(s):  
Electrospray Mass Spectrometry ◽  
Solid Phase Synthesis ◽  
Solid Phase ◽  
Chemical Properties ◽  
Oligonucleotide Synthesis ◽  
Phase Synthesis ◽  
Controlled Pore Glass ◽  
Product Identification ◽  
Pore Glass ◽  
And Chemical Properties

Polystyrene-polyethylene glycol (PS-PEG) tentacle polymers with loadings of up to 60/<μmol/g were used for standard oligonucleotide synthesis. As these resins are easy to handle and stable under reaction and cleavage conditions they may be used alternatively to controlled pore glass (CPG) as the most commonly used solid support for oligonucleotide synthesis. However, structural and chemical properties of the PS-PEG resins require modified conditions to guarantee syntheses with high coupling efficiencies. Oligonucleotides (ODN ) of various sequences and lengths have successfully been synthesized using HPLC and capillary electrophoresis (CE) for purity control. Additionally, electrospray mass spectrometry (ES-MS) was used for product identification.

Dansylated Aminopropyl Controlled Pore Glass:  A Model for Silica−Liquid Solvation+

Langmuir ◽  
2004 ◽  
Vol 20 (24) ◽  
pp. 10507-10516 ◽  
Author(s):  
Phillip M. Page ◽  
Chase A. Munson ◽  
Frank V. Bright
Keyword(s):  
Controlled Pore Glass ◽  
Pore Glass
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