Expression of Protein Complex Comprising the Human Prorenin and (Pro)Renin Receptor in Silkworm Larvae Using Bombyx mori Nucleopolyhedrovirus (BmNPV) Bacmids for Improving Biological Function
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Three forms of recombinant protein complexes comprising the human prorenin (hPro) and (pro)renin receptor (hPRR) (hPRR/prorenin) were successfully expressed in the silkworm larvae using Bombyx mori nucleopolyhedrovirus (BmNPV) bacmids. They were localized in the fat body cells and formed a prorenin-bound hPRR complex. The expressed levels of hPro and hPRR were similar judging from Western blotting. The hPRR/prorenin complex containing 40 μg of hPRR (yield, 43%) and 30 μg of hPro (yield, 34%) was purified from 15 silkworm larvae by a series of purification using anti-FLAG and Strep-Tactin affinity chromatography. The renin activity of the purified hPRR/prorenin complex was 3.8-fold that of the mixture of hPRR and hPro expressed individually in vitro judging from the renin assay. These results show that the unstable transmembrane protein, hPRR, was coexpressed stably with ligand, hPro, and formed a stable protein, hPRR/prorenin complex that showed a high catalytic active form.
KeywordsSilkworm Bombyx mori nucleopolyhedrovirus Bacmid (Pro)renin receptor Prorenin/(pro)renin receptor complex
- 7.Nguyen, G., Delarue, F., Burckle, C., Bouzhir, L., Giller, T., & Sraer, J. -D. (2002). Pivotal role of the renin/prorenin receptor in angiotensin II production and cellular responses to renin. The Journal of Clinical Investigation, 109, 1417–1427.Google Scholar
- 10.Ichihara, A., Hayashi, M., Kaneshiro, Y., Suzuki, F., Nakagawa, T., Tada, Y., et al. (2004). Inhibition of diabetic nephropathy by a decoy peptide corresponding to the “handle” region for nonproteolytic activation of prorenin. The Journal of Clinical Investigation, 114, 1128–1135.Google Scholar
- 15.Batenburg, W. W., Krop, M., Garrelds, I. M., de Vries, R., de Bruin, R. J., Burcklé, C. A., et al. (2007). Prorenin is the endogenous agonist of the (pro)renin receptor. Binding kinetics of renin and prorenin in rat vascular smooth muscle cells overexpressing the human (pro)renin receptor. Journal of Hypertension, 25, 2441–2453.CrossRefGoogle Scholar
- 18.Du, D., Kato, T., Suzuki F., & Park, E. Y. (2009). Binding affinity of full-length and extracellular domains of recombinant human (pro)renin receptors to human renin when expressed in the fat body and hemolymph of silkworm larvae. Journal of Bioscience and Bioengineering, in press.Google Scholar