Department of Biological Sciences, Bauru School of Dentistry, University of São Paulo, Bauru, São Paulo, Brazil.
Department of Genetics and Evolution, São Carlos Federal University, São Carlos, Brazil.
J Dent. 2022 Jan;116:103874. doi: 10.1016/j.jdent.2021.103874. Epub 2021 Nov 16.
To study the proteomic alterations in the initial AEP after rinsing with CaneCPI-5, StN15 or Hb or their combination.
In five crossover phases, after prophylaxis, 10 volunteers in 5 consecutive days, rinsed (10 mL, 1 min) with the following solutions: deionized water (HO- negative control- 1), 0.1 mg/mL CaneCPI-5 (2), 1.88×105 M StN15 (3), 1.0 mg/mL Hb (4) or their combination (5). The AEP formed after 3 min was collected with electrode filter papers soaked in 3% citric acid. After protein extraction, samples were analyzed by quantitative shotgun label-free proteomics.
Rinsing with the proteins/peptide increased the amounts of proteins in the AEP. The total numbers of proteins identified after rinsing with CaneCPI-5, StN15, Hb or their combination versus water, were 131, 167, 148 and 142, respectively. The treatment with the proteins/peptide or their combination increased proteins that bind calcium, phosphate and interact with distinct proteins, as well as proteins with antimicrobial and acid-resistant properties, such as, Cornifin-B (7.7, 12.6, and 4.3-fold for CaneCPI-5, StN15 and Hb, respectively), isoforms of Cystatin (2.2-2.4-fold for CaneCPI-5 and StN15), Proline-rich-protein 4 (4.3-fold; StN15), Histatin-1 (2.8-fold; StN15) and Hemoglobin (7.7-25-fold for Hb and Combination). Immunoglobulin, Keratin and Histone were exclusively identified upon treatment with the proteins/peptide, alone or combined.
Rinsing with proteins/peptide, alone or combined, increased protective proteins in the initial AEP.
Our results suggest that rinsing with the proteins/peptide or their combination increases the proteins capable of enhancing the protective function of the basal layer of AEP.
研究用 CaneCPI-5、StN15 或 Hb 或其组合冲洗后初始 AEP 中的蛋白质组变化。
在五个交叉阶段，在预防措施后，连续 5 天每天 10 名志愿者，用以下溶液冲洗（10 mL，1 分钟）：去离子水（HO-阴性对照-1）、0.1mg/mL CaneCPI-5（2）、1.88×105M StN15（3）、1.0mg/mL Hb（4）或其组合（5）。用浸泡在 3%柠檬酸中的电极滤纸收集 3 分钟后形成的 AEP。提取蛋白质后，通过定量鸟枪法无标记蛋白质组学进行分析。
用蛋白质/肽冲洗会增加 AEP 中的蛋白质数量。用 CaneCPI-5、StN15、Hb 或其组合冲洗后鉴定的总蛋白质数分别为 131、167、148 和 142，而用水冲洗则分别为 131、167、148 和 142。用蛋白质/肽或其组合处理会增加与钙、磷酸盐结合并与不同蛋白质相互作用的蛋白质，以及具有抗菌和耐酸特性的蛋白质，例如，角蛋白素-B（CaneCPI-5、StN15 和 Hb 分别为 7.7、12.6 和 4.3 倍）、半胱氨酸蛋白酶抑制剂同工型（CaneCPI-5 和 StN15 为 2.2-2.4 倍）、富含脯氨酸蛋白 4（StN15 为 4.3 倍）、组氨酸-1（StN15 为 2.8 倍）和血红蛋白（Hb 和组合分别为 7.7-25 倍）。免疫球蛋白、角蛋白和组蛋白仅在单独或联合使用蛋白质/肽处理时被鉴定。
单独或联合使用蛋白质/肽冲洗会增加初始 AEP 中的保护性蛋白质。
我们的结果表明，用蛋白质/肽或其组合冲洗会增加增强 AEP 基层保护功能的蛋白质。
