EVALUATION OF ESTROGENIC ACTIVITY OF PARABENS AND THEIR CHLORINATED DERIVATIVES BY USING THE YEAST TWO-HYBRID ASSAY AND THE ENZYME-LINKED IMMUNOSORBENT ASSAY

2009 ◽  
Vol 28 (1) ◽  
pp. 204 ◽  
Author(s):  
Masanori Terasaki ◽  
Ryo Kamata ◽  
Fujio Shiraishi ◽  
Masakazu Makino
Keyword(s):  
Immunosorbent Assay ◽  
Estrogenic Activity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Yeast Two Hybrid ◽  
Two Hybrid

Determination of estrogenic activity in landfill leachate by simplified yeast two-hybrid assay

2002 ◽  
Vol 4 (6) ◽  
pp. 1040-1046 ◽  
Author(s):  
Yasunori Kawagoshi ◽  
Yukiko Tsukagoshi ◽  
Isao Fukunaga
Keyword(s):  
Landfill Leachate ◽  
Estrogenic Activity ◽  
Yeast Two Hybrid ◽  
Two Hybrid

Microbial degradation of estrogens using activated sludge and night soil-composting microorganisms

2004 ◽  
Vol 50 (8) ◽  
pp. 153-159 ◽  
Author(s):  
J.H. Shi ◽  
Y. Suzuki ◽  
S. Nakai ◽  
M. Hosomi
Keyword(s):  
Activated Sludge ◽  
Microbial Degradation ◽  
Estrogenic Activity ◽  
Degradation Products ◽  
Order Reaction ◽  
Yeast Two Hybrid ◽  
First Order ◽  
Natural Estrogens ◽  
17Β Estradiol ◽  
Two Hybrid

In order to investigate the potential for microbial degradation of estrogens, and the products formed, activated sludge collected from Korea (ASK) and night soil-composting microorganisms (NSCM) were used to degrade estrogens. Results showed that both ASK and NSCM degraded almost 100% of the natural estrogens estrone (E1), 17β-estradiol (E2), and estriol (E3) from initial concentrations of 20-25 mg/L, while synthetic estrogen, ethynylestradiol (EE2), was not degraded. Analysis of degradation products of E2 by using HPLC-ECD and a consecutive first-order reaction calculation confirmed that E2 was sequentially degraded to E1, which was further degraded to other unknown compounds by ASK and NSCM. We then used the yeast two-hybrid assay to show that the unknown degradation products did not appear to possess estrogenic activity when E1, E2 or E3 were degraded to below the detection limit after 14 days of incubation, indicating that ASK and NSCM not only degrade natural estrogens, but also remove their estrogenic activities.

scholarly journals Genetic Analysis of theEscherichia coliFtsZ·ZipA Interaction in the Yeast Two-hybrid System

2001 ◽  
Vol 276 (15) ◽  
pp. 11980-11987 ◽  
Author(s):  
Steven A. Haney ◽  
Elizabeth Glasfeld ◽  
Cynthia Hale ◽  
David Keeney ◽  
Zhizhen He ◽  
...  
Keyword(s):  
Hybrid System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Wild Type ◽  
Yeast Two Hybrid ◽  
Conserved Sequence ◽  
Yeast Two Hybrid System ◽  
C Terminus ◽  
Two Hybrid

The recruitment of ZipA to the septum by FtsZ is an early, essential step in cell division inEscherichia coli. We have used polymerase chain reaction-mediated random mutagenesis in the yeast two-hybrid system to analyze this interaction and have identified residues within a highly conserved sequence at the C terminus of FtsZ as the ZipA binding site. A search for suppressors of a mutation that causes a loss of interaction (ftsZD373G) identified eight different changes at two residues within this sequence.In vitro, wild type FtsZ interacted with ZipA with a high affinity in an enzyme-linked immunosorbent assay, whereas FtsZD373Gfailed to interact. Two mutant proteins examined restored this interaction significantly.In vivo, the alleles tested are significantly more toxic than the wild typeftsZand cannot complement a deletion. We have shown that a fusion, which encodes the last 70 residues of FtsZ in the two-hybrid system, is sufficient for the interaction with FtsA and ZipA. However, when the wild type sequence is compared with one that encodes FtsZD373G, no interaction was seen with either protein. Mutations surrounding Asp-373 differentially affected the interactions of FtsZ with ZipA and FtsA, indicating that these proteins bind the C terminus of FtsZ differently.

scholarly journals Concentration and Inhibitor Removal Methods for Determination of Estrogenic Activity in River Water using Yeast Two-Hybrid Test

2004 ◽  
Vol 27 (11) ◽  
pp. 679-684 ◽  
Author(s):  
Masahiro HOSOKAWA ◽  
Rui LIU ◽  
Takashi KAMEYA ◽  
Takashi KUBO ◽  
Kohei URANO
Keyword(s):  
River Water ◽  
Estrogenic Activity ◽  
Yeast Two Hybrid ◽  
Hybrid Test ◽  
Two Hybrid

scholarly journals Estrogenic Activity of Branched 4-Nonylphenol Isomers Examined by Yeast Two-Hybrid Assay

2006 ◽  
Vol 52 (2) ◽  
pp. 132-141 ◽  
Author(s):  
Hirotaka Shioji ◽  
Shinji Tsunoi ◽  
Yosuke Kobayashi ◽  
Tatsushi Shigemori ◽  
Michihiko Ike ◽  
...  
Keyword(s):  
Estrogenic Activity ◽  
Yeast Two Hybrid ◽  
Two Hybrid

scholarly journals Estrogenic Activity of Coffee Constituents

Nutrients ◽  
2019 ◽  
Vol 11 (6) ◽  
pp. 1401 ◽  
Author(s):  
Ryoiti Kiyama
Keyword(s):  
Cell Signaling ◽  
Binding Assay ◽  
Estrogenic Activity ◽  
Yeast Two Hybrid ◽  
Nitrogenous Compounds ◽  
Protein Assay ◽  
Gene Assay ◽  
Bone Protection ◽  
Signal Crosstalk ◽  
Two Hybrid

Here, the constituents of coffee with estrogenic activity are summarized by a comprehensive literature search, and their mechanisms of action for their physiological effects are discussed at the molecular and cellular levels. The estrogenic activity of coffee constituents, such as acids, caramelized products, carbohydrates, lignin, minerals, nitrogenous compounds, oil (lipids), and others, such as volatile compounds, was first evaluated by activity assays, such as animal tests, cell assay, ligand-binding assay, protein assay, reporter-gene assay, transcription assay, and yeast two-hybrid assay. Second, the health benefits associated with the estrogenic coffee constituents, such as bone protection, cancer treatment/prevention, cardioprotection, neuroprotection, and the improvement of menopausal syndromes, were summarized, including their potential therapeutic/clinical applications. Inconsistent results regarding mixed estrogenic/anti-estrogenic/non-estrogenic or biphasic activity, and unbeneficial effects associated with the constituents, such as endocrine disruption, increase the complexity of the effects of estrogenic coffee constituents. However, as the increase of the knowledge about estrogenic cell signaling, such as the types of specific signaling pathways, selective modulations of cell signaling, signal crosstalk, and intercellular/intracellular networks, pathway-based assessment will become a more realistic means in the future to more reliably evaluate the beneficial applications of estrogenic coffee constituents.

scholarly journals The Multimerization of Hantavirus Nucleocapsid Protein Depends on Type-Specific Epitopes

2003 ◽  
Vol 77 (2) ◽  
pp. 943-952 ◽  
Author(s):  
Kumiko Yoshimatsu ◽  
Byoung-Hee Lee ◽  
Koichi Araki ◽  
Masami Morimatsu ◽  
Michiko Ogino ◽  
...  
Keyword(s):  
Amino Acids ◽  
Nucleocapsid Protein ◽  
Mammalian Cells ◽  
Competitive Elisa ◽  
Acid Number ◽  
Hantaan Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Yeast Two Hybrid ◽  
Seoul Virus ◽  
Two Hybrid

ABSTRACT Multimerization of the Hantaan virus nucleocapsid protein (NP) in Hantaan virus-infected Vero E6 cells was observed in a competitive enzyme-linked immunosorbent assay (ELISA). Recombinant and truncated NPs of Hantaan, Seoul, and Dobrava viruses lacking the N-terminal 49 amino acids were also detected as multimers. Although truncated NPs of Hantaan virus lacking the N-terminal 154 amino acids existed as a monomer, those of Seoul and Dobrava formed multimers. The multimerized truncated NP antigens of Seoul and Dobrava viruses could detect serotype-specific antibodies, whereas the monomeric truncated NP antigen of Hantaan virus lacking the N-terminal 154 amino acids could not, suggesting that a hantavirus serotype-specific epitope on the NP results in multimerization. The NP-NP interaction was also detected by using a yeast two-hybrid assay. Two regions, amino acids 100 to 125 (region 1) and amino acids 404 to 429 (region 2), were essential for the NP-NP interaction in yeast. The NP of Seoul virus in which the tryptophan at amino acid number 119 was replaced by alanine (W119A mutation) did not multimerize in the yeast two-hybrid assay, indicating that tryptophan 119 in region 1 is important for the NP-NP interaction in yeast. However, W119A mutants expressed in mammalian cells were detected as the multimer by using competitive ELISA. Similarly, the truncated NP of Seoul virus expressing amino acids 155 to 429 showed a homologous interaction in a competitive ELISA but not in the yeast two-hybrid assay, indicating that the C-terminal region is important for the multimerization detected by competitive ELISA. Combined, the results indicate that several steps and regions are involved in multimerization of hantavirus NP.

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