ORAL MUCOSAL PELLICLE - ADSORPTION AND TRANSPEPTIDATION OF SALIVARY COMPONENTS TO BUCCAL EPITHELIAL-CELLS
ORAL MUCOSAL PELLICLE - ADSORPTION AND TRANSPEPTIDATION OF SALIVARY COMPONENTS TO BUCCAL EPITHELIAL-CELLS
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DOI：
10.1042/bj2610887
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发表时间：
1989-08-01
影响因子：
4.1
通讯作者：
LEVINE, MJ
中科院分区：
文献类型：
--
作者：
BRADWAY, SD;BERGEY, EJ;LEVINE, MJ
本研究旨在探讨唾液分子与人颊上皮细胞相互作用的机制。利用人腮腺唾液抗血清，对上皮细胞1.5% SDS提取物进行电泳转移分析，检测筛选出的唾液成分。将这些细胞及其无细胞水萃取物与125- i标记的腮腺唾液孵育后，形成了一种碘化的高分子质量复合物，这种复合物在单独的125- i标记的生理盐水中不存在。这种复合物的形成是时间依赖性的，用EGTA、碘乙酰胺、n -乙基马来酰亚胺处理口腔上皮细胞或其无细胞提取物或在100℃下加热15分钟可以抑制这种复合物的形成。上皮细胞也促进了[13C]腐胺掺入到高分子质量复合物中，这种复合物的形成被碘乙酰胺、未标记的腐胺和EGTA抑制。细胞提取物介导的单氨尸胺交联成。α。这种相互作用被碘乙酰胺抑制。用4% (w/v) SDS/10% (v/v) β -巯基乙醇彻底提取125i -唾液或[14C]腐胺处理的上皮细胞后，上皮细胞包膜恢复了大量的放射性。经腐胺、EGTA、碘乙酰胺预处理或100℃加热15 min，可抑制放射性向上皮细胞包膜的掺入。这些数据表明：(1)口腔粘膜膜是唾液对上皮细胞质膜及其相关细胞骨架的选择性吸附形成的；(2)吸附的唾液成分可以通过上皮转谷氨酰胺酶相互交联或与上皮细胞骨架交联。
The present investigation was carried out to examine the mechanism(s) whereby salivary molecules interact with human buccal epithelial cells. By utilizing antiserum against human parotid saliva, selected salivary components were detected by electrophoretic-transfer analysis of 1.5% SDS extracts of epithelial cells. Incubation of the cells and their aqueous cell-free extracts with 125-I-labelled parotid saliva resulted in the formation of an iodinated high-molecular-mass complex which was not present in 125I-labelled saline alone. Formation of this complex was time-dependent and was inhibited by treating the buccal epithelial cells or their cell-free extracts with EGTA, iodoacetamide, N-ethylmaleimide or by heating at 100.degree.C for 15 min. The epithelial cells also promoted incorporation of [13C]putrescine into high-molecular-mass complexes whose formation was inhibited by iodoacetamide, unlabelled putrescine and EGTA. Cell extracts mediated cross-linking of monodansylcadaverine into .alpha.-casein, and this interaction was inhibited by iodoacetamide. Significant amounts of radioactivity were recovered with the epithelial-cell envelopes after exhaustive extraction of 125I-saliva- or [14C]putrescine-treated epithelial cells with 4% (w/v) SDS/10% (v/v) .beta.-mercaptoethanol. The incorporation of radioactivity into epithelial-cell envelopes was inhibited by pretreatment of the cells with putrescine, EGTA, iodacetamide or heating at 100.degree.C for 15 min. These data suggest that: (1) oral mucosal pellicvle is formed by the selective adsorption of saliva to the epithelial-cell plasma membrane and its associated cytoskeleton; and (2) the adsorbed salivary components may be cross-linked to each other or the epithelial cytoskeleton by epithelial transglutaminases.