Laboratory of Biotechnology, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
Laboratory of Biotechnology, Graduate School of Integrated Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka 422-8529, Japan.
Int J Mol Sci. 2019 Aug 29;20(17):4228. doi: 10.3390/ijms20174228.
Protein conjugations at post-translational levels are known to be essential to protein stability and function. Recently, it has been proven that the split protein CnaB2 (SpyTag/SpyCatcher, ST/SC) from can induce covalent conjugation rapidly and efficiently under various conditions. The protein of interest fused with the split protein SC/ST could be assembled spontaneously. In light of this finding, we introduced the ST/SC protein coupling concept into the silkworm-bacmid protein expression system (SpyBEVS). As a proof of concept, we first examined and confirmed that a competent ligation occurred between ST/SC-fused protein partners in vitro in cultured silkworm cells and in vivo in silkworm larvae by co-infection of several recombinant baculoviruses. The protein conjugation could be also achieved sufficiently by a simple one-step mixture of purified ST/SC-tagged peptide-protein pairs in vitro. Given the flexibility and robustness of silkworm-BEVS, our results on SpyBEVS show an alternative method for enabling the production of protein decorations in vitro and inside of silkworms.
蛋白质在翻译后水平的缀合被认为对蛋白质稳定性和功能至关重要。最近已经证明，来自 的分裂蛋白 CnaB2（SpyTag/SpyCatcher，ST/SC）可以在各种条件下快速有效地诱导共价缀合。与分裂蛋白 SC/ST 融合的靶蛋白可以自发组装。有鉴于此，我们将 ST/SC 蛋白偶联概念引入了家蚕-杆状病毒蛋白表达系统（SpyBEVS）。作为概念验证，我们首先通过共感染几种重组杆状病毒，在培养的家蚕细胞中和家蚕幼虫体内证实了 ST/SC 融合蛋白伴侣之间在体外和体内发生了有效的连接。通过在体外纯化的 ST/SC 标记肽-蛋白对的简单一步混合物也可以充分实现蛋白质缀合。鉴于家蚕-BEVS 的灵活性和稳健性，我们在 SpyBEVS 上的结果展示了一种替代方法，可以在体外和家蚕体内实现蛋白质修饰的产生。
