Department of Dentistry, Pontifical Catholic University of Minas Gerais, Av. Dom José Gaspar, 500, Prédio 46, Bairro Coração Eucarístico, Belo Horizonte, Minas Gerais 30535-610, Brazil.
Clin Oral Investig. 2012 Jun;16(3):761-6. doi: 10.1007/s00784-011-0556-5. Epub 2011 May 3.
Since in a previous study we encountered a subject with an unusual split MG2 banding pattern, the aim of this study was to investigate the molecular basis of this observation. Submandibular/sublingual secretion was collected under resting and stimulated conditions and examined on Western blots probed with anti-MG2 antibodies or on gels stained with periodic acid-Schiff reagent. Genomic DNA was isolated and the N-, tandem repeat (TR), and C-terminal regions of MUC7 were amplified by PCR since MG2 is known to display a genetic polymorphism. Although the typical appearance of MG2 on blots and gels is a single 180 kDa band, salivary secretions from the subject exhibited doublet immunoreactive bands of approximately 180 and 125 kDa. Additionally, under resting conditions the 180 kDa band was predominant whereas upon stimulation the 125 kDa band became predominant. Genomic DNA analysis showed that MUC7 in the individual with split MG2 bands was not truncated and that the MUC7 genotype in this individual was (6/6) where both alleles encoded six TRs. The MG2 split banding pattern observed in this subject was not derived from proteolytic degradation of this salivary mucin in whole saliva or from genetic polymorphism. The expression of two isoforms of MG2 could in principle improve or reduce the activity of this key component of the oral host defense system.
由于在之前的研究中,我们遇到了一个具有不寻常的 MG2 条带分裂模式的研究对象,因此本研究旨在探究这种观察结果的分子基础。在休息和刺激条件下收集颌下/舌下分泌物,并在 Western blot 上用抗 MG2 抗体探测或在经过碘酸-Schiff 试剂染色的凝胶上进行检查。由于 MG2 已知存在遗传多态性,因此分离基因组 DNA 并通过 PCR 扩增 MUC7 的 N-、串联重复(TR)和 C-末端区域。尽管 MG2 在印迹和凝胶上的典型外观是单个 180 kDa 条带,但研究对象的唾液分泌物显示出约 180 和 125 kDa 的双免疫反应性条带。此外,在休息条件下,180 kDa 条带占优势,而在刺激下,125 kDa 条带成为优势。基因组 DNA 分析表明,具有 MG2 分裂条带的个体中的 MUC7 没有截断,并且该个体的 MUC7 基因型为 (6/6),其中两个等位基因编码六个 TR。在这个研究对象中观察到的 MG2 分裂带模式不是来源于整个唾液中这种唾液粘蛋白的蛋白水解降解,也不是来源于遗传多态性。MG2 的两种同工型的表达原则上可以改善或降低口腔防御系统这一关键成分的活性。