Identification of lysine decarboxylase as a mammalian cell growth inhibitor in Eikenella corrodens:: possible role in periodontal disease
Identification of lysine decarboxylase as a mammalian cell growth inhibitor in Eikenella corrodens:: possible role in periodontal disease
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DOI：
10.1006/mpat.2000.0421
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发表时间：
2001-04-01
影响因子：
3.8
通讯作者：
Pan, HQ
中科院分区：
文献类型：
--
作者：
Levine, M;Progulske-Fox, A;Pan, HQ
通过研究腐蚀艾肯氏菌（Eikenella corrodens）无细胞盐溶性提取物和菌斑对HeLa和HL 60细胞生长的抑制机制，探讨了炎症性牙周病的发病机制。先前的研究发现一种蛋白质（p80）可引起大肠杆菌的生长抑制。腐蚀提取物。通过二维SDS-PAGE纯化后，用蛋白酶lysC消化p80。获得氨基酸序列并反向翻译用作PCR引物。从大肠杆菌Sau 3A 1基因组文库中获得一个含赖氨酸脱羧酶基因的5840个核苷酸的序列。会腐蚀DNA赖氨酸脱羧酶活性在生理pH下存在。腐蚀含有p80的提取物，也存在于菌斑中。两种浸提液均通过转化为尸胺消耗细胞培养基中的赖氨酸而引起生长抑制。将赖氨酸或免疫山羊IgG添加到衍生自E.腐蚀赖氨酸脱羧酶，延迟赖氨酸消耗和生长抑制。在HL 60细胞培养基中，E-氨基己酸在低赖氨酸浓度下特异性地增强赖氨酸脱羧酶活性，并且还增强生长抑制。因此，赖氨酸脱羧酶如p80通过从哺乳动物细胞培养基中除去赖氨酸来抑制生长。赖氨酸脱羧酶活性在牙周病的微生物病因学的新作用进行了讨论。(C)北京：科学出版社.
The pathogenesis of inflammatory periodontal disease was studied by examining the mechanism of HeLa and HL60 cell growth inhibition by cell-free saline-soluble extracts of Eikenella corrodens and bacterial plaque. Previous studies identified a protein (p80) as causing growth inhibition by E. corrodens extracts. After purification by two-dimensional SDS-PAGE, p80 was digested with protease lysC. Amino acid sequences were obtained and backtranslated for use as PCR primers. A 5840 nucleotide sequence containing a lysine decarboxylase gene was obtained from a Sau3A1 genomic library of E. corrodens DNA. Lysine decarboxylase activity was present at physiologic pH in the E. corrodens extracts containing p80, and also in bacterial plaque. Both extracts caused growth inhibition by depleting lysine from cell culture media through conversion to cadaverine. Adding lysine, or immune goat IgG to a peptide derived from the active site sequence of E. corrodens lysine decarboxylase, retarded lysine depletion and growth inhibition. E-Amino caproic acid specifically enhanced lysine decarboxylase activity at the low lysine concentration in HL60 cell culture media, and also increased the growth inhibition. Thus, lysine decarboxylases such as p80 inhibit growth by removing lysine from mammalian cell culture media. A new role for lysine decarboxylase activity in the microbial aetiology of periodontal disease is discussed. (C) 2001 Academic Press.