Arginine Catabolism and Its Role in Pathogenecity by Periodontopathogenic Bacteria

精氨酸分解代谢及其在牙周病原菌致病性中的作用

• 批准号: 12470461
• 负责人: NAKAMURA Ryo
• 金额: $ 7.87万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470461/
• 关键词: Growth of Porphyromonas gingivalis; Arginine carboxypeptidase; Arginine deiminase; Energy production; Periodontopathogenicity; 遺伝子解析; ATP産生; トリプシン様酵素; アルギニンデイミナーゼ

Porphyromonas gingivalis predominantly consumes arginine in the culture medium, suggesting that this amino acid could be the energy source. In relation to the arginine comsuption, cell extracts of P.gingivalis clearly demonstrated enzyme activities for the arginine deiminase pathway and adenosine triphosphate production. The pathogenic properties of this bacterium have been studied extensively in relation to the proteolytic enzyme, especially a trypsin-like enzyme, which splits the bond at the carboxyl side of arginine containing peptide. To obtain free arginine from protein and /or peptide, we performed to elucidate presence and role the arginine carboxypeptidase which cleaves peptide bond at the amino side of arginine. Arginine carboxypeptidase was found in the culture medium and cells. The enzyme was isolated and purified from cytoplasm of P.gingivalis cells. SDS-PAGE of the enzyme revealed the presence of three major bands of 42, 33, and 32kDa, of which 30 amino acid sequences at NH_2-terminal were identical. The ORF, suspected from the nucleotide sequences corresponding to the N-terminal amino acids on the date bases containing unfinished P.gingivalis W83 genome, showed to include signature, suggesting a zinc carboxypeptidase. By Western blotting and immunomicroscopy, the enzyme was found to distribute widely in the cytoplasm and on the surface of the outer membrane of P.gingivalis cells. These results show that this enzyme may function to release arginine in collaboration with a trypsin-like enzyme, to obtain arginine from periodontal tissues in the deep anaerobic pockets during the growth of P.gingivalis. Consequently, these processes might result in the pathogenecity of this bacterium.

牙龈卟啉单胞菌主要消耗培养液中的精氨酸，提示该氨基酸可能是能量来源。在精氨酸消耗方面，牙龈假单胞菌的细胞提取物明显显示出精氨酸脱亚胺酶途径和三磷酸腺苷产生的酶活性。该细菌的致病特性已被广泛地研究与蛋白水解酶，特别是类胰酶，它裂解含精氨酸的多肽的羧基一侧的键。为了从蛋白质和/或多肽中获得游离精氨酸，我们进行了精氨酸羧基肽酶的存在和作用的研究，该酶能裂解精氨酸氨基一侧的肽键。培养上清液和细胞中均有精氨酸羧肽酶的存在。该酶是从牙龈假单胞菌细胞质中分离纯化出来的。该酶的SDS-PAGE显示有42、33和32 kDa三条主带，其中NH_2末端有30个氨基酸序列相同。从含有未完成的牙龈假单胞菌W83基因组的碱基上的N-末端氨基酸对应的核苷酸序列中怀疑ORF包括签名，表明是锌羧肽酶。通过Western blotting和免疫显微镜观察发现，该酶广泛分布于牙龈假单胞菌细胞的胞浆和外膜表面。这些结果表明，在牙龈假单胞菌的生长过程中，该酶可能与一种类胰酶协同释放精氨酸，从牙周组织中的厌氧深层获得精氨酸。因此，这些过程可能导致这种细菌的致病性。

项目成果

中村亮 他1名(分担執筆): "歯周病-新しい治療を求めて-、歯周病原細菌の熱ショック蛋白質"医療先端技術研究所. 536 (2000)

Ryo Nakamura 和另一位作者（共同作者）：“牙周疾病 - 寻找新的治疗方法 - 牙周病原菌的热休克蛋白” 医学先进技术研究所 536（2000）。

Kaname Masuda et al.: "Consumption of peptide-derived arginine by periodontopathogenic bacterium, Porphyromonas gingivallis"Anaerobe. 7. 209-217 (2001)

Kaname Masuda 等人：“牙周致病菌牙龈卟啉单胞菌消耗肽衍生的精氨酸”厌氧菌。

Sugiyama, A., et al.: "Activation of human gingival epithelial cells by cell-surface components of black pigmented bacteria : augmentation od production of interleukin-8, granulocytecolony-stimulating factor and granulocytemacrophage colony-stimulating fa

Sugiyama, A. 等人：“黑色素细菌的细胞表面成分激活人类牙龈上皮细胞：增加白细胞介素 8、粒细胞集落刺激因子和粒细胞巨噬细胞集落刺激因子的产生

Tanabe, S., et al.: "Helicobacter pylori and Campylobater rectus share a common antigen"Oral Microbiol. Immnol.. 18 (in press). (2003)

Tanabe, S. 等人：“幽门螺杆菌和直肠弯曲菌拥有共同的抗原”口腔微生物。

Hinode Daisuke et al.: "Antigenic properties of the GroEL-like protein of Campyrobacter rectus"Oral Microbiol. Immunol. 17. 16-21 (2002)

Hinode Daisuke 等人：“直肠弯曲杆菌 GroEL 样蛋白的抗原特性”口腔微生物。