Properties of alkaline phosphatase in the gingival crevicular fluid.

J R Kina, N Yoshida, M Goseki, S Sasaki, I Ishikawa
{"title":"Properties of alkaline phosphatase in the gingival crevicular fluid.","authors":"J R Kina,&nbsp;N Yoshida,&nbsp;M Goseki,&nbsp;S Sasaki,&nbsp;I Ishikawa","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>The isoenzymic properties of the alkaline phosphatase (ALP) of the gingival crevicular fluid (GCF) were investigated and compared with those in other cells, such as human polymorphonuclear leukocytes (PMNs), and human periodontal ligament cells (PDLs), and with those of three species of periodontopathic bacteria: Porphyromonas gingivalis 381 (P. gingivalis), Prevotella intermedia ATCC25611 (P. intermedia), and Capnocytophaga sputigena ATCC33123 (C. sputigena). The biochemical properties of the isoenzymes were analyzed by the following methods: enzyme assays, inhibition pattern using three chemical inhibitors, 4 to 20% gradient polyacrylamide gel electrophoresis, thermostability, immunological specificity, and phosphatidylinositol-specific phospholipase C (PI-PLC) treatment. The inhibition experiment showed that ALP of the PMNs and PDLs possessed almost the same enzymatic properties of tissue-nonspecific ALP (bone/liver/kidney; TNSALP), and the ALP of the three species of periodontopathic bacteria possessed specific properties that were different from those of TNSALP, intestinal, or placental ALP. The ALP of the GCF was only slightly susceptible to levamisole (1 mM), L-phenylalanine (20 mM), and SDS (1%). An electrophoresis thermostability test demonstrated that the enzyme activity of the GCF was separated into one or two bands. The main heat-labile slow band contained the phosphatidylinositol (PI)-moiety-anchored ALP and possessed immunological specificity against anti-bone type ALP. The minor fast band was heat stable and showed mobility similar to that in P. gingivalis. These results indicated that the ALP of the GCF consisted of several ALP isoenzyme types whose possible origins are considered to be derived from phosphatidylinositol (PI) anchored ALP and periodontopathic bacterial ALP.(ABSTRACT TRUNCATED AT 250 WORDS)</p>","PeriodicalId":22311,"journal":{"name":"The Bulletin of Tokyo Medical and Dental University","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1995-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"The Bulletin of Tokyo Medical and Dental University","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

The isoenzymic properties of the alkaline phosphatase (ALP) of the gingival crevicular fluid (GCF) were investigated and compared with those in other cells, such as human polymorphonuclear leukocytes (PMNs), and human periodontal ligament cells (PDLs), and with those of three species of periodontopathic bacteria: Porphyromonas gingivalis 381 (P. gingivalis), Prevotella intermedia ATCC25611 (P. intermedia), and Capnocytophaga sputigena ATCC33123 (C. sputigena). The biochemical properties of the isoenzymes were analyzed by the following methods: enzyme assays, inhibition pattern using three chemical inhibitors, 4 to 20% gradient polyacrylamide gel electrophoresis, thermostability, immunological specificity, and phosphatidylinositol-specific phospholipase C (PI-PLC) treatment. The inhibition experiment showed that ALP of the PMNs and PDLs possessed almost the same enzymatic properties of tissue-nonspecific ALP (bone/liver/kidney; TNSALP), and the ALP of the three species of periodontopathic bacteria possessed specific properties that were different from those of TNSALP, intestinal, or placental ALP. The ALP of the GCF was only slightly susceptible to levamisole (1 mM), L-phenylalanine (20 mM), and SDS (1%). An electrophoresis thermostability test demonstrated that the enzyme activity of the GCF was separated into one or two bands. The main heat-labile slow band contained the phosphatidylinositol (PI)-moiety-anchored ALP and possessed immunological specificity against anti-bone type ALP. The minor fast band was heat stable and showed mobility similar to that in P. gingivalis. These results indicated that the ALP of the GCF consisted of several ALP isoenzyme types whose possible origins are considered to be derived from phosphatidylinositol (PI) anchored ALP and periodontopathic bacterial ALP.(ABSTRACT TRUNCATED AT 250 WORDS)

龈沟液中碱性磷酸酶的性质。
研究了龈沟液(GCF)碱性磷酸酶(ALP)的同工酶性质,并与其他细胞(如人多形核白细胞(PMNs)和人牙周韧带细胞(pdl))以及牙龈卟啉单胞菌381 (P. gingivalis)、中间普氏菌ATCC25611 (P. intermedia)和痰胞噬菌ATCC33123 (C. sputigena)的同工酶性质进行了比较。采用酶学分析、三种化学抑制剂的抑制模式、4 ~ 20%梯度聚丙烯酰胺凝胶电泳、热稳定性、免疫特异性和磷脂酰肌醇特异性磷脂酶C (PI-PLC)处理等方法分析了同工酶的生化特性。抑制实验表明,pmn和pdl的ALP具有与组织非特异性ALP(骨/肝/肾;三种牙周病细菌的ALP具有不同于TNSALP、肠道或胎盘ALP的特定性质。GCF的ALP对左旋咪唑(1 mM)、l -苯丙氨酸(20 mM)和SDS(1%)仅轻微敏感。电泳热稳定性测试表明,GCF的酶活性分为一个或两个条带。主要的热不稳定慢带含有磷脂酰肌醇(PI)片段锚定的ALP,对抗骨型ALP具有免疫特异性。次要的快带热稳定,其流动性与牙龈假单胞菌相似。这些结果表明,GCF的ALP由几种ALP同工酶组成,其可能的来源被认为是磷脂酰肌醇(PI)锚定的ALP和牙周病细菌ALP。(摘要删节250字)
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信