Human gingival crevicular fluid contains MRP8 (S100A8) and MRP14 (S100A9), two calcium-binding proteins of the S100 family
Human gingival crevicular fluid contains MRP8 (S100A8) and MRP14 (S100A9), two calcium-binding proteins of the S100 family
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DOI：
10.1177/00220345000790020701
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发表时间：
2000-02-01
影响因子：
7.6
通讯作者：
Cimasoni, G
中科院分区：
文献类型：
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作者：
Kojima, T;Andersen, E;Cimasoni, G
人类牙龈沟液含有未知的蛋白质，可能在牙周病中起标记作用。因此，本研究通过二维电泳（2-D PACE）分析，确定了在人龈沟液（GCF）中发现但在血清中不存在的低分子量蛋白。收集牙周炎和健康受试者以及无牙和新生儿受试者的GCF、血清和全唾液。蛋白质样品通过二维聚丙烯酰胺凝胶电泳分离，用银染色，并与SWISS-2D PAGE数据库中的参考蛋白图进行比较。在牙周炎患者和健康人的GCF和唾液中，观察到4个优势的低分子质量（8 ~ 14 kDa）酸性斑点。它们在无牙和新生儿的血清中没有发现，在唾液中也不太明显。从n端氨基酸测序中，鉴定出8 kDa和6.5 ~ 7.0等电点的2个二维蛋白点均为MRP8 （S100A8）蛋白，属于S100钙结合蛋白家族成员。利用多肽质量指纹图谱和基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）鉴定了另外两个蛋白点，质量为14 kDa，等电点在5.5 ~ 6.0之间，为蛋白MRP14 (S100A9)，也属于S100家族。Western blot证实GCF中存在MRP8和MRP14单克隆抗体。在GCF中鉴定的MRP8和MRP14两种多肽代表了血清和GCF的2-D PAGE模式之间的主要差异，我们假设它们可能在牙龈沟中发挥重要作用，并可能代表牙周病的可能标记物。
Human gingival crevicular fluid contains unidentified proteins which might play a role as markers in periodontal diseases. Therefore, low-molecular-weight proteins found in human gingival crevicular fluid (GCF), but absent from serum, were identified in the present study by means of two-dimensional electrophoresis (2-D PACE) analysis. GCF, serum, and whole saliva were collected from periodontitis and healthy subjects, as well as from edentulous and newborn subjects. Protein samples were separated by two-dimensional polyacrylamide gel electrophoresis, stained with silver, and compared with reference protein maps in the SWISS-2D PAGE database. In GCF and saliva from periodontitis patients and healthy subjects, four dominant low-molecular-mass (from 8 to 14 kDa) acidic spots were observed. They were not found in serum and were less visible in saliva from edentulous and newborn subjects. From N-terminal amino acid sequencing, the two 2-D protein spots of 8 kDa and isoelectric points between 6.5 and 7.0 were both identified as protein MRP8 (S100A8), a member of the S100 family of calcium-binding proteins. Using peptide mass fingerprinting and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), we identified the other two protein spots, with mass of 14 kDa and isoelectric points between 5.5 and 6.0, as protein MRP14 (S100A9), also belonging to the S100 family. The presence of MRP8 and MRP14 in GCF was confirmed by Western blot, with monoclonal antibodies. The two polypeptides, MRP8 and MRP14, identified in GCF represent the major difference between the 2-D PAGE patterns of serum and GCF, and we hypothesize that they may play an important role in the gingival sulcus and could represent possible markers for periodontal diseases.