Iontcheva I, Oppenheim F G, Troxler R F
Department of Periodontology and Oral Biology, School of Dental Medicine, Boston, Massachusetts 02118, USA.
J Dent Res. 1997 Mar;76(3):734-43. doi: 10.1177/00220345970760030501.
Heterotypic complexes between the high-molecular-weight mucin MG1 and other salivary proteins in human submandibular/sublingual secretion (HSMSL) could have a significant impact on the biological properties of these proteins in oral fluids in both health and disease. We describe a mild procedure for isolation and purification of native MG1 by gel filtration chromatography on Sepharose CL-2B which does not involve dialysis, lyophilization, use of denaturing agents, or covalent modification. Western blots of native MG1 probed with antibodies against 8 different salivary proteins showed that complexing occurs between MG1 and salivary amylase, proline-rich proteins (PRPs), statherins, and histatins but not MG1, sIgA, secretory component, or cystatins. When native MG1 was placed in 4 M guanidine hydrochloride and chromatographed on Sepharose CL-4B, ELISA measurement of column fractions showed that amylase, PRPs, statherins, and histatins were released. Interestingly, gel filtration resolved the material which eluted into 4 or 5 distinct peaks, suggesting that the released entities were heterotypic complexes. From these studies, the occurrence of at least three different types of complexes between MG1 and other salivary proteins has been identified. Type 1 complexes are dissociated by SDS-PAGE and in 4 M guanidine hydrochloride. Type II complexes are not dissociated under these conditions. Type III complexes are dissociated during SDS-PAGE and by 4 M guanidine hydrochloride, but the released proteins appear to be complexes containing amylase, PRPs, statherins, and histatins. The possible functional role of heterotypic complexes between MG1 and other salivary proteins as a physiologic delivery system, a mechanism for protection against proteolysis, a repository for precursors of the acquired enamel pellicle, and a vehicle for modulation of the viscoelastic and rheological properties of saliva is discussed.
高分子量粘蛋白MG1与人下颌下/舌下分泌液(HSMSL)中的其他唾液蛋白之间的异型复合物可能会对这些蛋白在健康和疾病状态下口腔液体中的生物学特性产生重大影响。我们描述了一种通过在Sepharose CL - 2B上进行凝胶过滤色谱法分离和纯化天然MG1的温和方法,该方法不涉及透析、冻干、使用变性剂或共价修饰。用针对8种不同唾液蛋白的抗体对天然MG1进行的蛋白质免疫印迹显示,MG1与唾液淀粉酶、富含脯氨酸的蛋白质(PRPs)、磷蛋白和组蛋白之间发生了复合,但与MG1、分泌型IgA、分泌成分或胱抑素之间没有复合。当将天然MG1置于4M盐酸胍中并在Sepharose CL - 4B上进行色谱分析时,对柱级分的酶联免疫吸附测定显示淀粉酶、PRPs、磷蛋白和组蛋白被释放出来。有趣的是,凝胶过滤将洗脱物分离成4个或5个不同的峰,这表明释放出的实体是异型复合物。通过这些研究,已确定MG1与其他唾液蛋白之间至少存在三种不同类型的复合物。I型复合物可通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE)和在4M盐酸胍中解离。II型复合物在这些条件下不会解离。III型复合物在SDS - PAGE过程中以及在4M盐酸胍作用下会解离,但释放出的蛋白质似乎是含有淀粉酶、PRPs、磷蛋白和组蛋白的复合物。本文讨论了MG1与其他唾液蛋白之间的异型复合物作为一种生理传递系统、一种防止蛋白水解的机制、获得性釉质 pellicle前体的储存库以及调节唾液粘弹性和流变学特性的载体的可能功能作用。