A new highly specific and robust yeast androgen bioassay for the detection of agonist and antagonists

T.F.H. Bovee, J.R. Helsdingen, A.R.M. Hamers, M. van Duursen, M.W.F. Nielen, L.A.P. Hoogenboom

Research output: Contribution to journalArticleAcademicpeer-review

72 Citations (Scopus)

Abstract

Public concern about the presence of natural and anthropogenic compounds which affect human health by modulating normal endocrine functions is continuously growing. Fast and simple high-throughput screening methods for the detection of hormone activities are thus indispensable. During the last two decades, a panel of different in vitro assays has been developed, mainly for compounds with an estrogenic mode of action. Here we describe the development of an androgen transcription activation assay that is easy to use in routine screening. Recombinant yeast cells were constructed that express the human androgen receptor and yeast enhanced green fluorescent protein (yEGFP), the latter in response to androgens. Compared with other reporters, the yEGFP reporter protein is very convenient because it is directly measurable in intact living cells, i.e., cell wall disruption and the addition of a substrate are not needed. When yeast was exposed to 17 beta-testosterone, the concentration where half-maximal activation is reached (EC50) was 50 nM. The relative androgenic potencies, defined as the ratio between the EC50 of 17 beta-testosterone and the EC50 of the compound, of 5 alpha-dihydrotestosterone, methyltrienolone, and 17 beta-boldenone are 2.3, 1.4, and 0.15 respectively. The results presented in this paper demonstrate that this new yeast androgen bioassay is fast, sensitive, and very specific and also suited to detect compounds that have an antiandrogenic mode of action.
Original languageEnglish
Pages (from-to)1549-1558
Number of pages10
JournalAnalytical and Bioanalytical Chemistry
Volume389
Issue number5
DOIs
Publication statusPublished - 2007

Keywords

  • green fluorescent protein
  • in-vitro
  • estrogenic activity
  • recombinant assay
  • response element
  • receptor-binding
  • surface waters
  • transcription
  • progesterone
  • validation

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