LARGE-SCALE PURIFICATION AND CHARACTERIZATION OF THE MAJOR PHOSPHOPROTEINS AND MUCINS OF HUMAN SUBMANDIBULAR-SUBLINGUAL SALIVA
LARGE-SCALE PURIFICATION AND CHARACTERIZATION OF THE MAJOR PHOSPHOPROTEINS AND MUCINS OF HUMAN SUBMANDIBULAR-SUBLINGUAL SALIVA
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DOI：
10.1042/bj2800341
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发表时间：
1991-12-01
影响因子：
4.1
通讯作者：
LEVINE, MJ
中科院分区：
文献类型：
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作者：
RAMASUBBU, N;REDDY, MS;LEVINE, MJ
人类颌下舌下唾液（HSMSL）的主要成分是粘蛋白、淀粉酶、半胱氨酸蛋白酶抑制剂、富含脯氨酸的蛋白质和富酪蛋白。这些分子的结构功能研究受到从人类分泌物中分离出的少量纯化材料的阻碍。本研究描述了用于大规模制备许多这些分子的集成纯化方案。为了解离唾液分子中的部分异型复合物，HSMSL 最初通过使用 6 M-盐酸胍的凝胶过滤分为四个池。随后通过凝胶过滤和离子交换色谱对这四个池进行分级分离，从而纯化了高和低 M(r) 粘蛋白、中性和酸性半胱氨酸蛋白酶抑制剂、酸性和碱性富含脯氨酸的蛋白质和富酪蛋白。这些唾液分子的许多变体或亚型已被鉴定并进行生化表征。生化研究表明，低 M(r) 粘蛋白以两种亚型存在，其唾液酸与岩藻糖的比例不同。酸性半胱氨酸蛋白酶抑制剂 S 的三种亚型的特征在于其磷酸盐含量不同。鉴定出一种富含脯氨酸的碱性肽的两种亚型；较小的肽是缺失前七个氨基酸的截短形式。
The major components of human submandibular-sublingual saliva (HSMSL) are mucins, amylases, cystatins, proline-rich proteins and statherin. Structure-function studies of these molecules have been hampered by the small amounts of purified materials that can be isolated from human secretions. The present study describes an integrated purification protocol for the large-scale preparation of many of these molecules. To dissociate partially heterotypic complexes among salivary molecules, HSMSL was initially fractionated into four pools by gel filtration with 6 M-guanidine hydrochloride. Subsequent fractionation of these four pools by gel-filtration and ion-exchange chromatography resulted in the purification of high- and low-M(r) mucins, neutral and acidic cystatins, acidic and basic proline-rich proteins and statherin. Many variants or isoforms of these salivary molecules have been identified and biochemically characterized. Biochemical studies indicated that the low-M(r) mucin exists as two isoforms which vary in their sialic acid to fucose ratios. Three isoforms of acidic cystatin S were characterized which differ in their phosphate content. Two isoforms of a basic proline-rich peptide were identified; the smaller peptide was a truncated form missing the first seven amino acids.