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Production of two recombinant insulin-like growth factor binding protein-1 subtypes specific to salmonids
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| Title: | Production of two recombinant insulin-like growth factor binding protein-1 subtypes specific to salmonids |
| Authors: | Hasegawa, Ryuya Browse this author | | Miura, Takuto Browse this author | | Kaneko, Nobuto Browse this author | | Kizaki, Ryousuke Browse this author | | Oishi, Gakuto Browse this author | | Tanaka, Hanae Browse this author | | Sato, Moe Browse this author | | Shimizu, Munetaka Browse this author →KAKEN DB |
| Keywords: | Whole genome duplication | | Tetraploidy | | Pituitary cells | | Growth hormone release | | Subfunction partitioning |
| Issue Date: | 1-Dec-2020 |
| Publisher: | Elsevier |
| Journal Title: | General and Comparative Endocrinology |
| Volume: | 299 |
| Start Page: | 113606 |
| Publisher DOI: | 10.1016/j.ygcen.2020.113606 |
| Abstract: | Salmonids have four subtypes of insulin-like growth factor binding protein (IGFBP)-1, termed -1a1, -1a2, -1b1 and 1b2, owing to teleost- and a lineage-specific whole-genome duplications. We have previously produced recombinant proteins of masu salmon IGFBP-1a1 and -1b2 and conducted functional analysis. To further characterize salmonid-specific IGFBP-1s, we cloned cDNAs encoding mature proteins of IGFBP-1a2 and -1b1 from the liver of masu salmon (Oncorhynchus masou). IGFBP-1a2 and -1b1 shared a 56% amino acid sequence homology whereas their homologies with their counterparts (i.e. -1a1 and -1b2) were 77% and 82%, respectively. We next expressed recombinant masu salmon (rs) IGFBP-1a2 and -1b1 with fusion partners thioredoxin (Trx) and a His-tag using the pET-32a(+) vector system in Escherichia coli. Trx.His.rsIGFBP-1s were detected in the insoluble faction, solubilized in a buffer containing urea, and isolated by Ni-affinity chromatography. They were refolded by dialysis and cleaved from the fusion partners by enterokinase. rsIGFBP-1a2 and -1b1 were purified by reversed-phase high performance liquid chromatography. Purified rsIGFBP-1a2 and -1b1 had the ability to bind digoxigenin-labeled human IGF-I on ligand blotting. We then examined the effects of rsIGFBP-1a1, -1a2, -1b1 and -1b2 in combination with human IGF-I on growth hormone (GH) release from cultured pituitary cells of masu salmon. IGF-I alone reduced GH release while the addition of rsIGFBP-1a1, -1b1 or -1b2, but not rsIGFBP-1a2, diminished the suppressive effect of IGF-I. Addition of rsIGFBP-1s without IGF-I had no effect on GH release. These results show that rsIGFBP-1b1, along with rsIGFBP-1a1 and -1b2, inhibits IGF-I action on the pituitary in masu salmon. The lack of the effect by rsIGFBP-1a2 suggests that salmon IGFBP-1 subtypes underwent subfunction partitioning and have different degrees of IGF-inhibitory action. |
| Rights: | © 2020. This manuscript version is made available under the CC-BY-NC-ND 4.0 license | | http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/83525 |
| Appears in Collections: | 水産科学院・水産科学研究院 (Graduate School of Fisheries Sciences / Faculty of Fisheries Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 清水 宗敬
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