Identification of lysine decarboxylase as a mammalian cell growth inhibitor in Eikenella corrodens: possible role in periodontal disease.
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 Sau3 A1 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. epsilon-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.
Levine
M
M
Department of Biochemistry & Molecular Biology, University of Oklahoma Health Sciences Center, P.O. Box 26901, Oklahoma City, Oklahoma, 73190, USA. martin-levine@ouhsc.edu
Progulske-Fox
A
A
Denslow
N D
ND
Farmerie
W G
WG
Smith
D M
DM
Swearingen
W T
WT
Miller
F C
FC
Liang
Z
Z
Roe
B A
BA
Pan
H Q
HQ
D002262:Carboxy-Lyases / Q000378:metabolism / Q000494:pharmacology*; D002455:Cell Division; D003470:Culture Media; D016124:Eikenella corrodens / Q000201:enzymology* / Q000472:pathogenicity; D006131:Growth Inhibitors / Q000494:pharmacology; D018922:HL-60 Cells; D006367:HeLa Cells; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008239:Lysine / Q000378:metabolism; D010510:Periodontal Diseases / Q000276:immunology / Q000382:microbiology* / Q000628:therapy
D003470:Culture Media; D006131:Growth Inhibitors; D007074:Immunoglobulin G; D002262:Carboxy-Lyases; C021520:lysine decarboxylase; D008239:Lysine