Gazi M I, Cox S W, Clark D T, Eley B M
Department of Periodontology, King's College School of Medicine and Dentistry, Denmark Hill, London, U.K.
Arch Oral Biol. 1995 Aug;40(8):731-6. doi: 10.1016/0003-9969(95)00032-k.
Earlier work has shown that gingival crevicular fluid (GCF) contains dipeptidyl peptidase (DPP) activities that resemble those in host tissue. Here, further comparisons were made with enzymes from suspected periodontal pathogens. Gingival tissue and GCF were collected from patients with chronic periodontitis. DPP II and DPP IV fractions with acid and alkaline pH optima, respectively, were separated from crude tissue extracts by gel-filtration chromatography. Bacterial cell sonicates were prepared from broth cultures of reference strains. There was moderate to strong DPP activity with Capnocytophaga spp., Porphyromonas gingivalis and Prevotella spp., very weak activity with Treponema denticola and no detectable activity with Actinobacillus actinomycetemcomitans or Fusobacterium nucleatum. Banding patterns in GCF, tissue and bacterial samples were compared on substrate-impregnated overlay membranes applied to isoelectric focusing gels. In gels washed with acid buffer, GCF had bands corresponding to tissue DPP II. Use of an alkaline washing buffer showed GCF activity which closely matched tissue DPP IV that had been pretreated with neuraminidase, an enzyme found by others in the gingival crevice. P. Gingivalis gave multiple bands and several of these had counterparts in GCF. The apparent presence in GCF of the DPP from P. gingivalis is consistent with the association of this organism with destructive periodontitis.
早期研究表明，龈沟液（GCF）中含有与宿主组织中相似的二肽基肽酶（DPP）活性。在此，对来自疑似牙周病原体的酶进行了进一步比较。从慢性牙周炎患者中收集牙龈组织和龈沟液。通过凝胶过滤色谱法从粗组织提取物中分离出分别具有酸性和碱性最适pH值的DPP II和DPP IV组分。从参考菌株的肉汤培养物中制备细菌细胞超声裂解物。二氧化碳嗜纤维菌属、牙龈卟啉单胞菌和普雷沃菌属具有中度至强的DPP活性，具核梭杆菌活性非常弱，而伴放线放线杆菌或具核梭杆菌未检测到活性。在应用于等电聚焦凝胶的底物浸渍覆盖膜上比较龈沟液、组织和细菌样品中的条带模式。在用酸性缓冲液洗涤的凝胶中，龈沟液具有与组织DPP II相对应的条带。使用碱性洗涤缓冲液显示龈沟液活性与用神经氨酸酶预处理的组织DPP IV紧密匹配，神经氨酸酶是其他人在龈沟中发现的一种酶。牙龈卟啉单胞菌产生多条条带，其中几条在龈沟液中有对应条带。龈沟液中明显存在来自牙龈卟啉单胞菌的DPP，这与该微生物与破坏性牙周炎的关联一致。
