Analogues of Human Parathyroid Hormone (1–31)NH2: Further Evaluation of the Effect of Conformational Constraint on Biological Activity

SUMMARY Human parathyroid hormone extracted from adenomata was assayed biologically by the release of calcium from mouse calvaria in organ culture and immunologically by immunoassay. The human hormone gave a logdose-response curve for calcium release and assayed at 12 units/mg in terms of the M.R.C. Bovine Research Standard A. Reduction with hot cysteine hydrochloride increased the biological activity of human hormone threefold, and the bovine hormone 4½-fold. An increase in immunological activity could only be shown with bovine hormone, and the purer the preparation the less was the activation of both biological and immunological potencies. Oxidation with hydrogen peroxide destroyed the biological activity of the bovine hormone but some residual immunological activity of the bovine and human hormones could always be detected. Back reduction with hot cysteine hydrochloride restored the biological and immunological activities of the bovine hormone and the immunological activity of the human hormone but never to the levels of the untreated hormones. It is suggested that these changes in potency can be induced by changes in the chemistry of the molecule rather than in its metabolism and that the immunological differences between bovine and human hormone represent minor differences in the amino acid sequence of these two hormones.

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Synthesis of human parathyroid-hormone-related protein(1-141) in Saccharomyces cerevisiae. A correct amino-terminal processing vital for the hormone's biological activity is obtained by an ubiquitin fusion protein approach

European Journal of Biochemistry ◽

10.1111/j.1432-1033.1993.tb17804.x ◽

1993 ◽

Vol 213 (1) ◽

pp. 648-648

Author(s):

Edith RIAN ◽

Rune JEMTLAND ◽

Ole K. OLSTAD ◽

Jan O. GORDELADZE ◽

Kaare M. GAUTVIK

Keyword(s):

Saccharomyces Cerevisiae ◽

Biological Activity ◽

Parathyroid Hormone ◽

Fusion Protein ◽

Related Protein ◽

Human Parathyroid Hormone ◽

Amino Terminal ◽

Parathyroid Hormone Related Protein

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Large-scale preparation and biological activity of recombinant human parathyroid hormone

Journal of Biotechnology ◽

10.1016/0168-1656(95)00002-8 ◽

1995 ◽

Vol 39 (2) ◽

pp. 129-136 ◽

Cited By ~ 3

Author(s):

J. Paulsen ◽

D. Ochs ◽

M. Harder ◽

C. Duvos ◽

H. Mayer ◽

...

Keyword(s):

Biological Activity ◽

Parathyroid Hormone ◽

Large Scale ◽

Human Parathyroid Hormone ◽

Large Scale Preparation ◽

Scale Preparation

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Comparative in vitro Biological Activity of 1–34 N-Terminal Synthetic Fragments of Human Parathyroid Hormone on Bovine and Porcine Kidney Membranes

Endocrine Research Communications ◽

10.3109/07435807609056898 ◽

1976 ◽

Vol 3 (3-4) ◽

pp. 167-186 ◽

Cited By ~ 4

Author(s):

C. A. Bader ◽

J. D. Monet ◽

P. Rivaille ◽

C. M. Gaubert ◽

M. S. Moukhtar ◽

...

Keyword(s):

Biological Activity ◽

Parathyroid Hormone ◽

Porcine Kidney ◽

Human Parathyroid Hormone

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Extraction and purification of recombinant intact human Parathyroid Hormone (hPTH) from bacterial cell

Journal of Biomedical Engineering and Informatics ◽

10.5430/jbei.v3n2p1 ◽

2017 ◽

Vol 3 (2) ◽

pp. 1 ◽

Cited By ~ 1

Author(s):

Adnan E. Al-badran ◽

Rafeef Amer Abdul-jabbar

Keyword(s):

Biological Activity ◽

Parathyroid Hormone ◽

Bacterial Cell ◽

Biologically Active ◽

Human Parathyroid Hormone ◽

Hormone Production ◽

Rp Hplc ◽

Bioactivity Assay ◽

Significant Difference ◽

Mcf 7

Objective: The intact human Parathyroid hormone (hPTH) is one of the biopharmaceutical drug produced by biotechnology, this hormone can be provided in a good amount using simple batch culture, and therefore the main purpose of this study was the production of purified bioactive intact hPTH.Methods: A cloning BL21(DE3) strain (which already cloned in our Lab. by synthetic human Parathyroid hormone (shPTH) gene) was grown in LB medium and the cloning gene expression was induced by addition of IPTG to the medium. The recombinant fused protein was purified from the bacterial cell lysate by affinity chromatography and underwent proteolysis by Enterokinase enzyme to obtain the target hPTH, and it's further purified by DEAE and SP Sepharose ion exchange chromatography. RP-HPLC analysis and biological activity on the MCF-7 breast cancer cells were done for both the standard and produced hPTH. Furthermore, the haemolysis ability of the latter was tested on the human RBC.Results: 225ng/ml of hPTH was produced after SP chromatography which detected by specific PTH ELISA kit, standard and produced hPTH showed the same peaks in the same retention time when analyzed by HPLC. The produced hPTH haemolysis assay showed the ability of the produced hPTH to partially haemolyze the RBC in a concentration above 200µg/ml. The bioactivity assay for the produced and standard rhPTH demonstrated that both compounds had a biological activity on the MCF-7 cells with IC50 of about 84.4 and 75.7 µg/ml for standard and produced hPTH respectively, and no significant difference was detected between them.Conclusions: Via suitable purification processes, the biologically active hPTH with structure similar to the standard ones as detected by RP-HPLC and bioactivity assay, can be obtained by using already cloned isolate with shPTH gene for hormone production. The hormone showed haemolysis effect on the RBC similar to the normal secreted hPTH.

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