Structural analysis of chicken oviduct progesterone receptor using monoclonal antibodies to the subunit B protein

Biochemistry ◽  
1984 ◽  
Vol 23 (19) ◽  
pp. 4427-4435 ◽  
Author(s):  
Dean P. Edwards ◽  
Nancy L. Weigel ◽  
William T. Schrader ◽  
Bert W. O'Malley ◽  
William L. McGuire
Keyword(s):  
Monoclonal Antibodies ◽  
Structural Analysis ◽  
Progesterone Receptor ◽  
Chicken Oviduct ◽  
Subunit B

scholarly journals Phosphorylation in vivo of chicken oviduct progesterone receptor.

1982 ◽  
Vol 257 (23) ◽  
pp. 14226-14230
Author(s):  
J J Dougherty ◽  
R K Puri ◽  
D O Toft
Keyword(s):  
Progesterone Receptor ◽  
Chicken Oviduct

Development and evaluation of an immunoenzymometric assay for the chicken progesterone receptor

1991 ◽  
Vol 129 (2) ◽  
pp. 189-196 ◽  
Author(s):  
M. K. Bläuer ◽  
P. J. Tuohimaa ◽  
P. J. Vilja
Keyword(s):  
Monoclonal Antibodies ◽  
Small Intestine ◽  
Horseradish Peroxidase ◽  
Progesterone Receptor ◽  
Ligand Binding ◽  
Binding Assay ◽  
Ligand Binding Assay ◽  
Coefficients Of Variation ◽  
First Time ◽  
Determination Range

ABSTRACT A specific and sensitive immunoenzymometric assay (IEMA) was developed for measuring the quantity of chicken progesterone receptor (PR) in tissue cytosol. The assay uses two monoclonal antibodies to the PR. One is used to capture the PR. The second (labelled with biotin) reacts first with the captured receptor and subsequently with avidin-labelled horseradish peroxidase to provide an enzymatic end-point. The method has a determination range from 0·3 to 60 pmol/l. Intra- and interassay coefficients of variation were 3·7% and 9·0% respectively. The assay can be performed with equal results as a rapid (3 h) or an overnight procedure. The IEMA is convenient, especially for signal measurement and the calculation of results. No ultracentrifugation of samples is needed, since the IEMA can be performed on low-speed cytosol samples. Assay results correlated well (r = 0·927) with those obtained by the conventional ligand-binding assay used in our laboratory. Similar results were obtained with the IEMA and the ligand-binding assay after exposure of cytosol samples to increased temperatures: at 20 °C the PR remained stable for the 4-h period examined, whereas at 37 °C almost complete degradation of the PR was observed in 30 min. Being more than 100 times as sensitive as the ligand-binding assay, the IEMA enabled the quantification of PR for the first time in such tissues as the bursa and small intestine even of immature animals. Journal of Endocrinology (1991) 129, 189–196

scholarly journals Monoclonal antibody to chicken oviduct progesterone receptor.

1983 ◽  
Vol 80 (10) ◽  
pp. 2854-2858 ◽  
Author(s):  
C. Radanyi ◽  
I. Joab ◽  
J. M. Renoir ◽  
H. Richard-Foy ◽  
E. E. Baulieu
Keyword(s):  
Monoclonal Antibody ◽  
Progesterone Receptor ◽  
Chicken Oviduct

Characterization of a novel 23-kilodalton protein of unactive progesterone receptor complexes

1994 ◽  
Vol 14 (3) ◽  
pp. 1956-1963
Author(s):  
J L Johnson ◽  
T G Beito ◽  
C J Krco ◽  
D O Toft
Keyword(s):  
Progesterone Receptor ◽  
Cdna Clone ◽  
Peptide Fragments ◽  
Receptor Complexes ◽  
Human Cdna ◽  
Partial Clone ◽  
Chicken Oviduct ◽  
Brain Cdna Library ◽  
Carboxy Terminal ◽  
And Function

Immunoprecipitation of unactivated avian progesterone receptor results in the copurification of hsp90, hsp70, and three additional proteins, p54, p50, and p23. p23 is also present in immunoaffinity-purified hsp90 complexes along with hsp70 and another protein, p60. Antibody and cDNA probes for p23 were prepared in an effort to elucidate the significance and function of this protein. Antibodies to p23 detect similar levels of p23 in all tissues tested and cross-react with a protein of the same size in mice, rabbits, guinea pigs, humans, and Saccharomyces cerevisiae, indicating that p23 is a conserved protein of broad tissue distribution. These antibodies were used to screen a chicken brain cDNA library, resulting in the isolation of a 468-bp partial cDNA clone encoding a sequence containing four sequences corresponding to peptide fragments isolated from chicken p23. This partial clone was subsequently used to isolate a full-length human cDNA clone. The human cDNA encodes a protein of 160 amino acids that does not show homology to previously identified proteins. The chicken and human cDNAs are 88% identical at the DNA level and 96.3% identical at the protein level. p23 is a highly acidic phosphoprotein with an aspartic acid-rich carboxy-terminal domain. Bacterially overexpressed human p23 was used to raise several monoclonal antibodies to p23. These antibodies specifically immunoprecipitate p23 in complex with hsp90 in all tissues tested and can be used to immunoaffinity isolate progesterone receptor complexes from chicken oviduct cytosol.

scholarly journals Structural Analysis of Monoclonal Antibodies by Ultrahigh Resolution MALDI In-Source Decay FT-ICR Mass Spectrometry

2018 ◽  
Vol 91 (3) ◽  
pp. 2079-2085 ◽  
Author(s):  
Yuri E.M. van der Burgt ◽  
David P. A. Kilgour ◽  
Yury O. Tsybin ◽  
Kristina Srzentić ◽  
Luca Fornelli ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibodies ◽  
Structural Analysis ◽  
Ultrahigh Resolution ◽  
Ft Icr

scholarly journals Immunogenetic and structural analysis of a class of HCV broadly neutralizing antibodies and their precursors

2018 ◽  
Vol 115 (29) ◽  
pp. 7569-7574 ◽  
Author(s):  
Fernando Aleman ◽  
Netanel Tzarum ◽  
Leopold Kong ◽  
Kenna Nagy ◽  
Jiang Zhu ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Structural Analysis ◽  
Neutralizing Antibodies ◽  
Antigenic Site ◽  
The Other ◽  
Maturation Process ◽  
Broadly Neutralizing Antibodies ◽  
Vaccine Candidates ◽  
Rational Vaccine Design ◽  
Hairpin Conformation

Elicitation of broadly neutralizing antibodies (bnAbs) is a leading strategy in rational vaccine design against antigenically diverse pathogens. Here, we studied a panel of monoclonal antibodies (mAbs) from mice immunized with the hepatitis C virus (HCV) envelope glycoproteins E1E2. Six of the mAbs recognize the conserved E2 antigenic site 412–423 (AS412) and cross-neutralize diverse HCV genotypes. Immunogenetic and structural analysis revealed that the antibodies originated from two different germline (GL) precursors and bind AS412 in a β-hairpin conformation. Intriguingly, the anti-HCV activity of one antibody lineage is associated with maturation of the light chain (LC), whereas the other lineage is dependent on heavy-chain (HC) maturation. Crystal structures of GL precursors of the LC-dependent lineage in complex with AS412 offer critical insights into the maturation process of bnAbs to HCV, providing a scientific foundation for utilizing the mouse model to study AS412-targeting vaccine candidates.

scholarly journals Progesterone receptor in the chick oviduct: an immunohistochemical study with antibodies to distinct receptor components.

1984 ◽  
Vol 99 (4) ◽  
pp. 1193-1201 ◽  
Author(s):  
J M Gasc ◽  
J M Renoir ◽  
C Radanyi ◽  
I Joab ◽  
P Tuohimaa ◽  
...  
Keyword(s):  
Progesterone Receptor ◽  
Polyclonal Antibodies ◽  
Smooth Muscles ◽  
Immunoperoxidase Technique ◽  
Cell Nuclei ◽  
Receptor Complexes ◽  
B Subunit ◽  
Chicken Oviduct ◽  
A Subunit ◽  
Immunohistochemical Studies

We performed immunohistochemical studies of chicken oviduct after different fixation procedures, by using antibodies against the progesterone receptor: polyclonal antibodies IgG-G3 against the "8S" form (an oligomere containing progesterone-binding and nonprogesterone-binding units), polyclonal antibodies IgG-RB against the progesterone-binding B subunit, and monoclonal BF4 against the non-progesterone-binding 90,000-mol-wt protein component. Chickens were immature animals with or without estrogen priming, and with or without progesterone treatment. The antibodies were revealed by means of an immunoperoxidase technique that used the avidin-biotin-peroxidase complex, and controls were performed by presaturation of antibodies with the purified 8S-progesterone receptor, the B subunit, and 90,000-mol-wt protein. The progesterone receptor was detected not only in well-characterized target tissues, i.e., in glands and luminal epithelium, but also in stromal cells (some displayed the strongest reaction), in mesothelium, and in fibers of smooth muscles. Only in cell nuclei, whether or not the animals received an injection of progesterone was an antigen revealed corresponding to the B subunit (and/or to the A subunit, because there is immunoreactivity of IgG-RB with both hormone-binding subunits A and B). The 90,000-mol-wt protein was revealed in both cytoplasm and nuclei. These immunohistological data suggest that the concept of steroid action that necessarily involves the original formation of the hormone-receptor complexes in the cytoplasm before translocation to the nucleus, may have to be revised.

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