Binding Affinity of Full-length and Extracellular Domains of Recombinant Human (Pro)renin Receptor to Human Renin When Expressed in the Fat Body and Hemolymph of Silkworm Larvae.
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概要
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Transmembrane domains of some receptors have been found to be very important in the process of constitutive oligomerization, and in the stability and functioning of the receptor. In this study, full-length of human (pro) renin receptor (hPRR) and hPRR lacking cytoplasmic domain (hPRR-ΔCD) were expressed in fat body of silkworm larvae, and the extracellular domain of hPRR (hPRR-ΔTMΔCD) in hemolymph. Three forms of hPRR were used for investigation of the interaction between receptor and ligand using surface plasmon resonance (SPR). As a result, the cytoplasmic domain was not an essential requirement for binding affinity, but the transmembrane domain of hPRR was indispensable in the formation of functional hPRR. The dissociation equilibrium constants (K_D) of purified hPRR and hPRR-ΔCD were estimated to be 46nM and 330nM, respectively. No evidence of binding by the extracellular domain of hPRR located in hemolymph was found. However, the solubilized microsomal fraction of the extracellular domain of hPRR expressed in the fat body showed specific affinity, but lost its binding affinity after purification. Its binding affinity was recovered by mixing microsomal fraction of mock-injected fat body to the purified extracellular domain. It is probable that an artificial transmembrane domain stabilizes the extracellular domain of hPRR and native conformation may be structurally recovered. To our knowledge, these are the first findings describing the interaction of transmembrane and extracellular domains of hPRR with ligand and this may help towards the understanding of binding affinity of hPRR to ligand.
著者
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Park Enoch
Laboratory Of Biotechnology Department Of Applied Biological Chemistry Shizuoka University:integrate
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Du Dongning
Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technolog
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Kato Tatsuya
Laboratory of Biotechnology, Faculty of Agriculture, Shizuoka University
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Suzuki Fumiaki
Laboratory of Animal Biochemistry, Faculty of Applied Biological Sciences, Gifu University
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Park Enoch
Laboratory Of Biotechnology Integrated Bioscience Section Graduate School Of Science And Technology
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Park Enoch
Laboratory Of Biotechnology Department Of Applied Biological Chemistry Faculty Of Agriculture Shizuo
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Du Dongning
Laboratory Of Biotechnology Integrated Bioscience Section Graduate School Of Science And Technology
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Kato Tatsuya
Laboratory Of Biotechnology Faculty Of Agriculture Shizuoka University
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Suzuki Fumiaki
Laboratory Of Animal Biochemistry Faculty Of Applied Biological Sciences Gifu University
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Kato Tatsuya
Laboratory Of Biotechnology Department Of Applied Biological Chemistry Shizuoka University
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Kato Tatsuya
Laboratory Of Biotechnology Department Of Applied Biological Chemistry Faculty Of Agriculture Shizuo
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