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Conversion of thyrotropin heterodimer to a biologically active single-chain
F A Fares1, S Yamabe, D Ben-Menahem
1Department of Biochemistry, Carmel Medical Center, and the Rappaport Family Institute for Research in the Medical Sciences, Faculty of Medicine, Technion-Israel Institute of Technology, Haifa. biochem@actcom.co.il
Endocrinology
|May 16, 1998
Summary
Researchers created a single-chain version of thyroid-stimulating hormone (TSH) for improved study. This novel approach enhances TSH secretion while maintaining receptor binding and bioactivity, aiding future structure-function research.
Area of Science:
- Endocrinology
- Molecular Biology
- Glycoprotein Hormone Research
Background:
- Thyroid-stimulating hormone (TSH) and gonadotropins (FSH, LH, CG) are heterodimeric glycoprotein hormones.
- Proper assembly of alpha and beta subunits is crucial for hormone function, but mutagenesis studies can hinder this process.
Purpose of the Study:
- To develop a biologically active single-chain human TSH (hTSH) to overcome challenges in traditional structure-function studies.
- To investigate the effect of a carboxyl terminal peptide (CTP) linker on single-chain hTSH secretion and activity.
Main Methods:
- Genetically engineered a single-chain hTSH by fusing the alpha and beta subunits, with and without a hCGbeta CTP linker.
- Expressed wild-type hTSH and single-chain variants in Chinese hamster ovary (CHO) cells.
- Assessed hormone secretion, receptor binding, and in vitro bioactivity.
Main Results:
- Single-chain hTSH was efficiently secreted from CHO cells, with a 3-fold increase compared to wild-type hTSH.
- Secretion of the single-chain hTSH was dramatically enhanced when using the CTP linker.
- Receptor binding and in vitro bioactivity of the single-chain variants remained comparable to wild-type hTSH.
Conclusions:
- The single-chain approach is a viable strategy for producing biologically active TSH.
- This method significantly improves TSH secretion, offering a valuable tool for future structure-function relationship investigations.
- The use of a CTP linker further optimizes the secretion of single-chain TSH.