Sorsa T, Ding Y L, Ingman T, Salo T, Westerlund U, Haapasalo M, Tschesche H, Konttinen Y T
Department of Periodontology, University of Helsinki, Finland.
J Clin Periodontol. 1995 Sep;22(9):709-17. doi: 10.1111/j.1600-051x.1995.tb00831.x.
Dental plaque is the major aetiological factor in periodontal diseases and contains several proteolytic enzymes. The origin of these proteinases is, however, poorly studied. This study was undertaken to characterize collagenase present in dental plaque of adult periodontitis patients. Vertebrate-type rather than bacterial-derived collagenase activity was detected in extracts of both supra- and subgingival dental plaque extracts of adult periodontitis patients. Dental plaque collagenase was found to exist predominantly in autoactive form. Dental plaque collagenase from periodontally healthy individuals existed in latent from. Latent dental plaque collagenase from periodontitis lesions could be activated by a 95 kD chymotrypsin-like proteinase from Treponema denticola and human leukocyte cathepsin G but not by human plasmin. Incubation of purified latent leukocyte collagenase with whole cells of Fusobacterium nucleatum, Eubacterium saburreum, Prevotella buccae and Porphyromonas gingivalis, however, did not result to the activation of the enzyme. Doxycycline in vitro inhibited dental plaque collagenase with an IC50-value of 20 microM. Dental plaque collagenase degraded more efficiently type I and II collagens than type III collagen. Western-blot analysis with specific anti-human neutrophil collagenase-antibody revealed that both in supra- and subgingival dental plaque extracts dental plaque collagenase had undergone proteolytic conversion from an 80 kD proform to a 58 kD active form which is associated with catalytic autoactivity as measured by functional collagenase assay. This reflects proteolytic activation of leukocyte collagenase in dental plaque probably by other proteases derived from potent periodontopathogenic bacteria such as T. denticola or other PMN proteases such as cathepsin G.(ABSTRACT TRUNCATED AT 250 WORDS)
牙菌斑是牙周疾病的主要病因，其中含有多种蛋白水解酶。然而，这些蛋白酶的来源却鲜有研究。本研究旨在鉴定成年牙周炎患者牙菌斑中的胶原酶特性。在成年牙周炎患者的龈上和龈下牙菌斑提取物中均检测到脊椎动物型而非细菌来源的胶原酶活性。发现牙菌斑胶原酶主要以自激活形式存在。牙周健康个体的牙菌斑胶原酶以潜伏形式存在。来自牙周炎病变的潜伏性牙菌斑胶原酶可被具核梭杆菌、粘性真杆菌、颊普氏菌和牙龈卟啉单胞菌的一种95kD类胰凝乳蛋白酶样蛋白酶以及人白细胞组织蛋白酶G激活，但不能被人纤溶酶激活。然而，将纯化的潜伏性白细胞胶原酶与具核梭杆菌、粘性真杆菌、颊普氏菌和牙龈卟啉单胞菌的全细胞一起孵育，并未导致该酶的激活。体外实验中，强力霉素抑制牙菌斑胶原酶，IC50值为20μM。牙菌斑胶原酶对I型和II型胶原的降解效率高于III型胶原。用特异性抗人中性粒细胞胶原酶抗体进行的蛋白质印迹分析表明，在龈上和龈下牙菌斑提取物中，牙菌斑胶原酶均经历了蛋白水解转化，从80kD的前体形式转变为58kD的活性形式，通过功能性胶原酶测定，该活性形式与催化自活性相关。这反映了牙菌斑中白细胞胶原酶可能被来自强效牙周病原菌（如齿垢密螺旋体）的其他蛋白酶或其他PMN蛋白酶（如组织蛋白酶G）进行蛋白水解激活。（摘要截短至250字）
