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The relationship between the connecting peptide of recombined single chain insulin and its biological function

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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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Authors and Affiliations

  1. State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 200031, Shanghai, China

    Yiding Huang, Zhenhe Liang & Youmin Feng

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  1. Yiding Huang
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  2. Zhenhe Liang
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  3. Youmin Feng
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Correspondence to Youmin Feng.

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Huang, Y., Liang, Z. & Feng, Y. The relationship between the connecting peptide of recombined single chain insulin and its biological function. Sci. China Ser. C.-Life Sci. 44, 593–600 (2001). https://doi.org/10.1007/BF02879353

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  • DOI: https://doi.org/10.1007/BF02879353

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