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Construction and Comparable Activity of A Novel hGHRH Analogs, Pro-hGHRH(1-44)-Gly-Gly-Cys+ |
Song-Shan TANG |
Guangdong Pharmaceutical University |
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Abstract AIM: Growth hormone releasing hormone (GHRH) is one of the hypothalamus hormones. Because of its potential applications in agriculture and human medicine, an analog with high GHRH activity and prolonged half-life has been looked for. Methods: By using the fusion protein expression protocol with unique acid labile linker Asp-Pro, the Pro-hGHRH(1-44)-Gly-Gly-Cys peptide was obtained. The peptide was purified to homogeneity by means of cell disruption, washing of inclusion body, ethanol fraction precipitation, acid hydrolysis, SP-Sephadex C-25 and Sephadex G-10 column chromatography. Results: The peptide molecular weight of 5,373 Da measured by EIS-MS is coincident with the actual values and its purity was determined by SDS-PAGE. The peptide at 0.1-10 μg/ml increased GH releases both from human pituitary and from rat pituitary. The rat pituitary GH-releasing activity increased in a dose-dependent manner. Compared with the standard hGHRH(1-40), the two GHRH analogs had higher rGH-releasing activities. The standard hGHRH(1-40) at 0.01-2.0 μg/ml showed no measured rat GH release. The activity comparisons showed that the rat pituitary GH-releasing activity of the Pro-GHRH(1-44)Gly-Gly-Cys peptide was not more than that of the Pro-hGHRH(1-44) peptide, whereas the human pituitary GH-releasing activity of the Pro-GHRH(1-44)-Gly-Gly-Cys peptide was far less than that of the Pro-hGHRH(1-44) peptide although there was not significant between them. Conclusion: The activity results showed that they had good GH-releasing activity, function-selecting and species specificity.
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Received: 07 May 2009
Published: 10 May 2010
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Corresponding Authors:
Song-Shan TANG
E-mail: songstang@gmail.com
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