Full-length chicken parathyroid hormone: Biosynthesis in escherichia coli and analysis of biologic activity

S. K. Lim, T. Gardella, A. Thompson, J. Rosenberg, H. Keutmann, J. Potts, H. Kronenberg, S. Nussbaum

Research output: Contribution to journalArticle

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Abstract

Chicken parathyroid hormone (cPTH) has been reported to stimulate adrenal steroidogenesis and to have unusual potency on traditional PTH target tissues. To evaluate these properties, chicken PTH-(1-88) has been expressed in Escherichia coli using a plasmid encoding a fusion protein which links together growth hormone, a factor Xa recognition site, and chicken PTH-(1-88). The growth hormone-cPTH fusion protein required the presence of 0.02% sodium dodecyl sulfate to remain in solution and be cleaved by factor Xa. The high performance liquid chromatography-purified recombinant cPTH-(1-88) and chemically synthesized cPTH-(1-34) had similar potency in rat osteosarcoma (ROS 17/2.8) cells, opossum kidney (OK) cells, and dispersed primary chicken kidney cells. The biologic potencies of cPTH-(1-34) and cPTH-(1-88) in radioreceptor binding and cAMP generation in both bone- and kidney-derived cell lines were less than those of human (h)PTH-(1-34). In dispersed chicken kidney cells, cAMP production by cPTH-(1-34) and cPTH-(1-88) was similar to that stimulated by human PTH-(1-34). No stimulation of steroidogenesis could be detected when recombinant chicken PTH-(1-88) was added to dispersed chicken adrenal cells. The biologic activity of recombinant chicken PTH-(1-88) purified from E. coli was comparable with that of chicken PTH-(1-88) expressed by mammalian COS cells. Thus, the full-length chicken PTH did not exhibit enhanced potency, when compared with human PTH in ROS 17/2.8, OK cell lines, and dispersed chicken kidney cells and did not demonstrate the novel steroidogenic action previously reported in adrenal cells. The successful production of chicken PTH-(1-88) will enhance our understanding of the structure-activity relationships for PTH, particularly the sequence-dependent metabolism of the hormone.

Original languageEnglish
Pages (from-to)3709-3714
Number of pages6
JournalJournal of Biological Chemistry
Volume266
Issue number6
Publication statusPublished - 1991 Jul 17

Fingerprint

Biosynthesis
Parathyroid Hormone
Escherichia coli
Chickens
Growth Hormone
Kidney
Fusion reactions
Cells
Factor Xa
Opossums
High performance liquid chromatography
Metabolism
Sodium Dodecyl Sulfate
Rats
Bone
Proteins
Plasmids
Hormones
Tissue
Cell Line

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lim, S. K., Gardella, T., Thompson, A., Rosenberg, J., Keutmann, H., Potts, J., ... Nussbaum, S. (1991). Full-length chicken parathyroid hormone: Biosynthesis in escherichia coli and analysis of biologic activity. Journal of Biological Chemistry, 266(6), 3709-3714.
Lim, S. K. ; Gardella, T. ; Thompson, A. ; Rosenberg, J. ; Keutmann, H. ; Potts, J. ; Kronenberg, H. ; Nussbaum, S. / Full-length chicken parathyroid hormone : Biosynthesis in escherichia coli and analysis of biologic activity. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 6. pp. 3709-3714.
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Lim, SK, Gardella, T, Thompson, A, Rosenberg, J, Keutmann, H, Potts, J, Kronenberg, H & Nussbaum, S 1991, 'Full-length chicken parathyroid hormone: Biosynthesis in escherichia coli and analysis of biologic activity', Journal of Biological Chemistry, vol. 266, no. 6, pp. 3709-3714.

Full-length chicken parathyroid hormone : Biosynthesis in escherichia coli and analysis of biologic activity. / Lim, S. K.; Gardella, T.; Thompson, A.; Rosenberg, J.; Keutmann, H.; Potts, J.; Kronenberg, H.; Nussbaum, S.

In: Journal of Biological Chemistry, Vol. 266, No. 6, 17.07.1991, p. 3709-3714.

Research output: Contribution to journalArticle

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AU - Lim, S. K.

AU - Gardella, T.

AU - Thompson, A.

AU - Rosenberg, J.

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AU - Kronenberg, H.

AU - Nussbaum, S.

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Lim SK, Gardella T, Thompson A, Rosenberg J, Keutmann H, Potts J et al. Full-length chicken parathyroid hormone: Biosynthesis in escherichia coli and analysis of biologic activity. Journal of Biological Chemistry. 1991 Jul 17;266(6):3709-3714.