Interaction between human augmenter of liver regeneration and Na+-K+-ATPase in vitro |
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| Authors: | TONG Ming-hua KONG Xiang-ping LI Mai YANG Lian-ping YANG Ru-de |
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| Institution: | 1.Center of Liver Diseases of PLA, The 458th Hospital of PLA, Guangzhou 510602, China;2.Department of Biotechnology, South China University of Technology, Guangzhou 510641, China. E-mail:minghuat@tom.com |
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| Abstract: | AIM: To check the physical interaction between GST- Na+-K+-ATPase domain and recombinant human augmenter of liver regeneration (rhALR) by GST pull down assay. METHODS: With PCR and genetic recombinant techniques, the coding region of β subunit of Na+-K+-ATPase was cloned into expressing plasmid pGEX-4T and identified by endonuclease digestion and sequencing methods. Under the inducing of 0.1 mmol/L IPTG, the fusion protein GST- Na+-K+-ATPase domain was highly expressed by E.coli DH-5α. After hypersound quassating, the GST- Na+-K+-ATPase domain was purified by glutathione agarose beads and the physical interaction with rhALR was checked by GST pull down assay. RESULTS: Analysis by SDS-PAGE showed the rhALRs of monomer and dimmer in GST- Na+-K+-ATPase domain lane. The Western blotting of the GST-pull down assay showed the same results as well. CONCLUSION: The Na+-K+-ATPase domain is associated with rhALR specifically in vitro. |
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| Keywords: | Augmenter of liver regeneration Na+-K+-ATPase |
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