The relationship between the connecting peptide of recombined single chain insulin and its biological function |
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Authors: | Yiding Huang Zhenhe Liang and Youmin Feng |
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Institution: | State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China |
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Abstract: | To investigate the relationship between the biological activity of recombined single chain insulin and the length of the connecting
peptide, we designed and prepared three single chain insulin molecules, namely, PIP, A]5PIP and A]10PIP, by site-directed mutagenesis, in which B30 and A1 were linked through dipeptide A-K, heptapeptide A-A-A-A-A-A-K, and
dodecapeptide A-A-A-A-A-A-A-A-A-A-A-K, respectively. Their receptor binding capacities were 0.14%, 14.3% and 11.1% of that
of insulin respectively and theirin vivo biological activities were in consistence with their receptor binding capacity; whereas their growth promoting activities
were 17%, 116.3% and 38% of that of insulin. These results suggested the following conclusions. (i) The recombined single
chain insulin could also possess the same metabolic and mitogenic function as insulin. (ii) The receptor binding capacity
of recombined single chain insulin to insulin receptor was closely related to the length and amino acid composition of the
connecting peptide and could change from 0 to 100% of insulin depending on the different connecting peptides. This result
further illustrated the necessity of B chain C-terminus swaying away from A chain N-terminus when insulin binds to its receptor.
(iii) The mitogenic activity of recombined single chain insulin also depended on the length and the amino acid composition
of the connecting peptide and was higher than its metabolic activity. |
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Keywords: | insulin single chain insulin receptor binding growth promoting GR2H6 cell line |
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