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马基莓半胱氨酸蛋白酶抑制剂:重组表达、特性鉴定及其在保护牙齿牙本质和牙釉质中的应用

Maquiberry Cystatins: Recombinant Expression, Characterization, and Use to Protect Tooth Dentin and Enamel.

作者信息

de Souza Eduardo Pereira, Ferro Milene, Pelá Vinicius Taioqui, Fernanda-Carlos Thais, Borges Cecília Guimarães Giannico, Taira Even Akemi, Ventura Talita Mendes Oliveira, Arencibia Ariel Domingo, Buzalaf Marília Afonso Rabelo, Henrique-Silva Flávio

机构信息

Department of Genetics and Evolution, Federal University of São Carlos (UFSCar), São Carlos 13565-905, SP, Brazil.

Department of General and Applied Biology, Institute of Biosciences, São Paulo State University (UNESP), Rio Claro 13506-900, SP, Brazil.

出版信息

Biomedicines. 2023 May 4;11(5):1360. doi: 10.3390/biomedicines11051360.

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 (), 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, < 0.05), suggesting its potential usage in dental products.

摘要

植物半胱氨酸蛋白酶抑制剂是参与植物生理和防御作用的半胱氨酸蛋白酶的蛋白质竞争性抑制剂。有人提出将其作为治疗人类疾病的潜在疗法,因此在不同植物(如智利酒果)中寻找新型半胱氨酸蛋白酶抑制剂变体是有意义的。作为一个研究较少的物种,智利酒果蛋白质的生物技术潜力鲜为人知。在本研究中,我们使用下一代测序技术构建了智利酒果幼苗的转录组,从中发现了六个半胱氨酸蛋白酶抑制剂序列。其中五个被克隆并进行了重组表达。针对木瓜蛋白酶、人组织蛋白酶B和L进行了抑制试验。智利酒果半胱氨酸蛋白酶抑制剂可以以纳摩尔级抑制蛋白酶,但智利酒果半胱氨酸蛋白酶抑制剂4和5除外,它们以微摩尔级抑制组织蛋白酶B。这表明智利酒果半胱氨酸蛋白酶抑制剂在治疗人类疾病方面具有潜在用途。此外,由于我们之前证明了一种甘蔗来源的半胱氨酸蛋白酶抑制剂对保护牙釉质的功效,我们测试了智利酒果半胱氨酸蛋白酶抑制剂-3保护牙本质和牙釉质的能力。两者均受到该蛋白质的保护(通过单因素方差分析和Tukey多重比较检验,P<0.05),表明其在牙科产品中的潜在用途。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4bb8/10216612/81f4c624e107/biomedicines-11-01360-g001.jpg

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