PDF(Pubmed)
Abstract：
Phytocystatins are proteinaceous competitive inhibitors of cysteine peptidases involved in physiological and defensive roles in plants. Their application as potential therapeutics for human disorders has been suggested, and the hunt for novel cystatin variants in different plants, such as maqui (Aristotelia chilensis), is pertinent. Being an understudied species, the biotechnological potential of maqui proteins is little understood. In the present study, we constructed a transcriptome of maqui plantlets using next-generation sequencing, in which we found six cystatin sequences. Five of them were cloned and recombinantly expressed. Inhibition assays were performed against papain and human cathepsins B and L. Maquicystatins can inhibit the proteases in nanomolar order, except MaquiCPIs 4 and 5, which inhibit cathepsin B in micromolar order. This suggests maquicystatins\' potential use for treating human diseases. In addition, since we previously demonstrated the efficacy of a sugarcane-derived cystatin to protect dental enamel, we tested the ability of MaquiCPI-3 to protect both dentin and enamel. Both were protected by this protein (by One-way ANOVA and Tukey\'s Multiple Comparisons Test, p < 0.05), suggesting its potential usage in dental products.
摘要：
植物细胞抑制素是参与植物生理和防御作用的半胱氨酸肽酶的蛋白质竞争性抑制剂。它们作为人类疾病的潜在治疗方法已经被提出，以及在不同植物中寻找新的胱抑素变体，例如maqui（辣椒木兰），是相关的。作为一个研究不足的物种，对maqui蛋白的生物技术潜力知之甚少。在本研究中,我们使用下一代测序构建了一个maqui小植株的转录组，其中我们发现了六个胱抑素序列。其中5个被克隆并重组表达。对木瓜蛋白酶和人组织蛋白酶B和L进行抑制试验。Maquicystatin可以以纳摩尔顺序抑制蛋白酶，除了MaquiCPIs4和5，它们以微摩尔顺序抑制组织蛋白酶B。这表明maquicystatins可用于治疗人类疾病。此外,因为我们以前证明了甘蔗衍生的胱抑素保护牙釉质的功效，我们测试了MaquiCPI-3保护牙本质和牙釉质的能力.两者都受到这种蛋白质的保护(通过单向方差分析和Tukey的多重比较测试，p<0.05)，表明其在牙科产品中的潜在用途。
