Department of Parasitology School of Medicine, Pusan National University, Yangsan, Republic of Korea.
Department of Molecular Biology, College of Natural Sciences, Pusan National University, Busan, Republic of Korea.
PLoS Negl Trop Dis. 2018 Nov 1;12(11):e0006516. doi: 10.1371/journal.pntd.0006516. eCollection 2018 Nov.
In a previous study, we found that Trichinella spiralis muscle larva excretory and secretory proteins (ES-P) most likely activate collagen synthesis via TGF-β/Smad signaling, and this event could influence collagen capsule formation.
METHODOLOGY/PRINCIPAL FINDINGS: In order to identify the specific collagen inducing factor, ES-P was fractionated by a Superdex 200 10/300 GL column. We obtained three large fractions, F1, F2, and F3, but only F3 had collagen gene inducing ability. After immunoscreening, 10 collagen inducing factor candidates were identified. Among them, TS 15-1 and TS 15-2 were identical to the putative trypsin of T. spiralis. The deduced TS 15-1 (M.W. = 72 kDa) had two conserved catalytic motifs, an N-terminal Tryp_SPc domain (TS 15-1n) and a C-terminal Tryp_SPc domain (TS 15-1c). To determine their collagen inducing ability, recombinant proteins (rTS 15-1n and rTS 15-1c) were produced using the pET-28a expression system. TS 15-1 is highly expressed during the muscle larval stage and has strong antigenicity. We determined that rTS 15-1c could elevate collagen I via activation of the TGF-β1 signaling pathway in vitro and in vivo.
CONCLUSION/SIGNIFICANCE: In conclusion, we identified a host collagen inducing factor from T. spiralis ES-P using immunoscreening and demonstrated its molecular characteristics and functions.
在之前的研究中，我们发现旋毛虫肌肉幼虫排泄分泌蛋白（ES-P）很可能通过 TGF-β/Smad 信号通路激活胶原蛋白合成，这一事件可能影响胶原蛋白囊的形成。
方法/主要发现：为了鉴定特定的胶原蛋白诱导因子，我们通过 Superdex 200 10/300 GL 柱对 ES-P 进行了分级。我们得到了三个大的部分，F1、F2 和 F3，但只有 F3 具有胶原蛋白基因诱导能力。经过免疫筛选，我们鉴定出了 10 个胶原蛋白诱导因子候选物。其中，TS 15-1 和 TS 15-2 与旋毛虫的假定胰蛋白酶相同。推断的 TS 15-1（M.W. = 72 kDa）具有两个保守的催化基序，一个 N 端 Tryp_SPc 结构域（TS 15-1n）和一个 C 端 Tryp_SPc 结构域（TS 15-1c）。为了确定它们的胶原蛋白诱导能力，我们使用 pET-28a 表达系统生产了重组蛋白（rTS 15-1n 和 rTS 15-1c）。TS 15-1 在肌肉幼虫期高度表达，具有很强的抗原性。我们确定 rTS 15-1c 可以通过体外和体内激活 TGF-β1 信号通路来提高胶原蛋白 I 的水平。
总之，我们使用免疫筛选从旋毛虫 ES-P 中鉴定出一种宿主胶原蛋白诱导因子，并证明了其分子特征和功能。
