Bolscher J G, Groenink J, van der Kwaak J S, van den Keijbus P A, van 't Hof W, Veerman E C, Nieuw Amerongen A V
Academic Centre for Dentistry Amsterdam, Department of Oral Biochemistry, The Netherlands.
J Dent Res. 1999 Jul;78(7):1362-9. doi: 10.1177/00220345990780071101.
The large carbohydrate moiety of low-Mr salivary mucin MUC7 (originally referred to as MG2) is subject to variations. Biochemical analysis and quantification of MUC7 in saliva samples require recognition tools that are independent of the carbohydrate moiety. Therefore, we have evoked three antisera to synthetic peptides of MUC7. One of these (CpMG2), raised against the C-terminal peptide, recognized native MUC7 in saliva and was characterized further. Recognition of MUC7 by CpMG2 turned out to be specific, resistant to dissociating and reductive treatments, and independent of glycosylation differences, as indicated by Western analysis and ELISA. The antiserum could be used to monitor MUC7 during purification procedures. MUC7 was demonstrated in small volumes of saliva from all (sero)mucous glands, including the palate and lip. Analysis with antibodies and lectins indicated large variations in amount as well as in glycosylation of MUC7. An ELISA was developed to determine the relative quantity of MUC7 in the glandular salivas: mean values of approximately 220, 980, and 100 microg mucin per mL were found in submandibular, sublingual, and palatine saliva, respectively.
低分子量唾液粘蛋白MUC7(最初称为MG2)的大碳水化合物部分存在变异。唾液样本中MUC7的生化分析和定量需要独立于碳水化合物部分的识别工具。因此,我们制备了三种针对MUC7合成肽的抗血清。其中一种(CpMG2)是针对C末端肽产生的,可识别唾液中的天然MUC7,并对其进行了进一步表征。如Western分析和ELISA所示,CpMG2对MUC7的识别具有特异性,对解离和还原处理具有抗性,且与糖基化差异无关。该抗血清可用于在纯化过程中监测MUC7。在包括腭和唇在内的所有(浆液性)粘液腺的少量唾液中均检测到了MUC7。用抗体和凝集素进行的分析表明,MUC7的含量和糖基化存在很大差异。开发了一种ELISA方法来测定腺性唾液中MUC7的相对含量:在下颌下腺、舌下腺和腭腺唾液中,每毫升粘蛋白的平均值分别约为220、980和100微克。