scholarly journals Studies on sex-organ development. Isolation and characterization of an oestrogen receptor from chick Müllerian duct

1975 ◽  
Vol 150 (2) ◽  
pp. 183-190 ◽  
Author(s):  
C S Teng ◽  
C T Teng
Keyword(s):  
Chick Embryo ◽  
Isoelectric Focusing ◽  
Oestrogen Receptor ◽  
Gradient Analysis ◽  
Hormone Receptors ◽  
Mullerian Duct ◽  
Müllerian Duct ◽  
High Affinity ◽  
Isolation And Characterization

An oestradiol-binding macromolecule was observed in the left Müllerian duct of the 15-day female chick embryo. The embryonic receptor binds oestradiol with a high affinity and low capacity, having a Kd of 3.2 × 10(-9)M and a maximal number of sites of 5.45 fmol/10(6) cells in the left Müllerian duct. The receptor is protein in nature, as suggested by its susceptibility to proteolysis; in addition, it is organ- and steroid-specific. Judging by glycerol-gradient analysis, the hormone receptors in the cytosol are present in 8S and 4.5S forms, and the 8S form could be dissociated into a 4.5S form in the presence of 0.5M-KCl. A 4.5-6S receptor could be extracted from the nuclei. Under physiological salt conditions, the embryonic receptors bind to DNA-cellulose and can be eluted when the salt concentration is increased to 0.5M-KCl. Determination by isoelectric focusing indicates that the isoelectric point is 5.8 for the 8S and 6.9 for the 4.5S receptor.

scholarly journals Carbohydrate binding activities of Bradyrhizobium japonicum. II. Isolation and characterization of a galactose-specific lectin.

1990 ◽  
Vol 111 (4) ◽  
pp. 1639-1643 ◽  
Author(s):  
S C Ho ◽  
M Schindler ◽  
J L Wang
Keyword(s):  
Isoelectric Focusing ◽  
Bradyrhizobium Japonicum ◽  
Affinity Column ◽  
Carbohydrate Binding ◽  
Binding Affinities ◽  
Isolation And Characterization ◽  
Relative Binding ◽  
Mannose Derivatives ◽  
Derivatives Of

Extracts of Bradyrhizobium japonicum were fractionated on Sepharose columns covalently derivatized with lactose. Elution of the material that was specifically bound to the affinity column with lactose yielded a protein of Mr approximately 38,000. Isoelectric focusing of this sample yielded two spots with pI values of 6.4 and 6.8. This protein specifically bound to galactose-containing glycoconjugates, but did not bind either to glucose or mannose. Derivatives of galactose at the C-2 position showed much weaker binding; there was an 18-fold difference in the relative binding affinities of galactose versus N-acetyl-D-galactosamine. These results indicate that we have purified a newly identified carbohydrate-binding protein from Bradyrhizobium japonicum, that can exquisitely distinguish galactose from its derivatives at the C-2 position.

scholarly journals Isolation and characterization of human plasma α 1-proteinase inhibitor and a conformational study of its interaction with proteinases

1976 ◽  
Vol 157 (2) ◽  
pp. 339-351 ◽  
Author(s):  
J Saklatvala ◽  
G C Wood ◽  
D D White
Keyword(s):  
Human Plasma ◽  
Isoelectric Focusing ◽  
Proteinase Inhibitor ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sedimentation Equilibrium ◽  
Isolation And Characterization ◽  
Step Procedure ◽  
Helical Content

1. alpha 1-Proteinase inhibitor was isolated from human plasma by a five-step procedure. Isoelectric focusing showed that six components focused between pH4.85 and 4.95. 2. The mol.wt. of the inhibitor was 52000 by sedimentation equilibrium and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The amino acid and carbohydrate compositions of the inhibitor were also determined. 3. The far-u.v.c.d. (circular-dichroism) spectrum indicated that the inhibitor had about 36% alpha-helical content. 4. The loss of proteinase-inhibitory activity when the inhibitor was exposed to pH values less than 5.0 or greater than 10.5 was accompanied by small changes in the far-u.v.c.d. spectrum and large changes in the near-u.v.c.d. spectrum. The change at alkaline pH was associated with ionization of tyrosine residues. 5. Interaction of inhibitor with chymotrypsin caused perturbation of the c.d. spectrum and this was used to follow the interaction and show a 1:1 stoicheiometry. 6. C.d., electrophoresis and isoelectric focusing showed that the inhibitor-enzyme complex is degraded by free enzyme. 7. Parallel studies with trypsin indicated that it too forms a 1:1 complex with inhibitor and is degraded by excess of enzyme.

scholarly journals Isolation and characterization of membrane proteins responsible for attachment of polyribosomes to rough microsomal fraction of rat liver

1977 ◽  
Vol 164 (1) ◽  
pp. 53-66 ◽  
Author(s):  
S Fujita ◽  
F Ogata ◽  
J Nakamura ◽  
S Omata ◽  
H Sugano
Keyword(s):  
Rat Liver ◽  
Protein Fraction ◽  
Microsomal Fraction ◽  
Binding Capacity ◽  
Gel Filtration ◽  
High Affinity ◽  
Isolation And Characterization ◽  
Microsomal Membranes ◽  
Equilibrium Centrifugation

A protein fraction which has a high affinity for polyribosomes was isolated from rough microsomal membranes of rat liver. The mode of polyribosome binding to this fraction (R-fraction) was studied by using CsCl equilibrium centrifugation and compared with that for stripped rough microsomal membranes. The following were found. (1) The polyribosome-binding cpacity of the R-fraction was heat-labile and sensitive to trypsin, and was suppressed by increasing KCl concentration and addition of 0.1 mM-aurintricarboxylic acid. (2) Of the four subfractions obtained by gel filtration of the R-fraction on a Sephadex G-200, only the R1-fraction, eluted at the void volume, showed a high affinity for polyribosomes. The polyribosome-binding capacity of the R1-fraction decreased with time on storage at 4 degrees C. (3) The R1-fraction contained three major proteins with mol. wts. 108,000, 99,000 and 65,000.

scholarly journals TCR-based therapy for multiple myeloma and other B-cell malignancies targeting intracellular transcription factor BOB1

Blood ◽  
2017 ◽  
Vol 129 (10) ◽  
pp. 1284-1295 ◽  
Author(s):  
Lorenz Jahn ◽  
Pleun Hombrink ◽  
Renate S. Hagedoorn ◽  
Michel G. D. Kester ◽  
Dirk M. van der Steen ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Transcription Factor ◽  
B Cell ◽  
B Cell Malignancies ◽  
High Affinity ◽  
Isolation And Characterization ◽  
Key Points

Key Points Isolation and characterization of a high-affinity TCR targeting the intracellular B cell–specific transcription factor BOB1. T cells expressing a BOB1-specific TCR lysed and eradicated primary multiple myeloma and other B-cell malignancies in vitro and in vivo.

scholarly journals Isolation and characterization of high affinity and highly stable anti-Chikungunya virus antibodies using ALTHEA Gold Libraries™

2020 ◽  
Author(s):  
Martha Pedraza-Escalona ◽  
Omar Guzmán-Bringas ◽  
Ivan Arrieta-Oliva ◽  
Keyla Gómez-Castellano ◽  
Juana Salinas-Trujano ◽  
...  
Keyword(s):  
Chikungunya Virus ◽  
Dynamic Range ◽  
Sandwich Elisa ◽  
Diagnostic Methods ◽  
Diagnostic Tools ◽  
Chikv Infection ◽  
High Affinity ◽  
Isolation And Characterization ◽  
Sds Page

Abstract Background More than three million infections were attributed to Chikungunya virus (CHIKV) in the 2014–2016 outbreak in Mexico, Central and South America, with over 500 deaths directly or indirectly related to this viral disease. CHIKV outbreaks are recurrent and no vaccine nor approved therapeutics exist to prevent or treat CHIKV infection. Reliable and robust diagnostic methods are thus critical to control future CHIKV outbreaks. Direct CHIKV detection in serum samples via highly specific and high affinity anti-CHIKV antibodies has shown to be an early and effective clinical diagnosis. Methods Chikungunya virions isolated from serum of a patient in Veracruz, México, were purified and characterized via electron microscopy, SDS-PAGE and binding to diverse well-characterized anti-CHIKV monoclonal antibodies. UV-inactivated CHIKV particles were used as selector in a solid-phase panning coupled with ELISA-based screening and Next-Generation Sequencing to discover specific and high affinity anti-CHIKV antibodies from ALTHEA Gold Libraries™. Results The CHIKV isolate showed the typical morphology of the virus. Protein bands in the SDS-PAGE were consistent with the size of its capsid proteins. UV-inactivated CHIKV particles bound tightly the control antibodies. The lead antibodies here obtained showed high expression yield, monomeric content over 95% after a single-step Protein A purification, and importantly, a thermal stability above 75oC. Most of the antibodies recognized linear epitopes on E2, including the highest affinity antibody called C7. A sandwich ELISA implemented with C7 and a potent neutralizing antibody isolated elsewhere, also specific for E2 but recognizing a discontinuous epitope, showed a dynamic range of 0.2–40.0 µγ/mL of UV-inactivated CHIKV purified preparation. The number of CHIKV particles estimated based on the concentration of E2 in the extract suggested that the assay could detect clinically meaningful amounts of CHIKV in serum. Conclusions The newly discovered antibodies offer valuable tools for characterization of CHIKV isolates and development of robust diagnostic tools for CHIKV infection surveillance. Application of ALTHEA Gold Libraries™ in combination with viral particles other than CHIKV could expedite the discovery and development of antibodies for detection and control of emergent and quickly spreading viral outbreaks such as SARS-CoV-2 (COVID-19).

Isolation and characterization of a high-affinity ammonium transporter ApAMT1;1 in alligatorweed

2019 ◽  
Vol 89 (3) ◽  
pp. 321-330 ◽  
Author(s):  
Xiaotong Guo ◽  
Yuting Sheng ◽  
Shunying Yang ◽  
Lei Han ◽  
Yachao Gao ◽  
...  
Keyword(s):  
Ammonium Transporter ◽  
High Affinity ◽  
Isolation And Characterization
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