Hydrolysis of plasmalogen by phospholipase A1 from Streptomyces albidoflavus for early detection of dementia and arteriosclerosis.
OBJECTIVE
To obtain an ethanolamine plasmalogen (PlsEtn)-hydrolyzing enzyme and to develop an assay that would help determine PlsEtn concentrations in human serum as an indicator of Alzheimer-type dementia and of arteriosclerosis.


RESULTS
Phospholipase A1s, SaPLA1 and SvPLA1 from, respectively, Streptomyces albidoflavus NA297 and S. avermitilis JCM5070-but not phospholipase B from Streptomyces sp. NA684, PLA2-Nagase from S. avermitilis, PLA2IIL from S. violaceoruber nor LIPOMOD 699L (porcine phospholipase)-hydrolyzed choline plasmalogen (PlsCho) and PlsEtn (PlsCho preferred over PlsEtn). Using a combination of SaPLA1, lysoplasmalogen-specific phospholipase D (LyPls-PLD), with amine oxidase, an end-point assay was developed for measuring serum PlsEtn concentration. The standard curve, generated using various amounts of PlsEtn in this assay, was linear between 0 and 0.2 mM. PlsEtn concentrations in forty-seven serum samples, determined independently by this enzyme-based assay and (125)I-HPLC method, exhibited a linear relationship, indicating that the assay is suitable for fast and accurate measurement of serum PlsEtn concentration.


CONCLUSIONS
An assay, developed using SaPLA1, LyPls-PLD, and AOX, selectively measured PlsEtn levels in blood samples. This assay could be a useful diagnostic tool for early stage detection of diseases such as Alzheimer-type dementia and arteriosclerosis.
Asahi Kasei Pharma Corp., 632-1 Mifuku, Izunokuni, Shizuoka, 410-2321, Japan. sakasegawa.sb@om.asahi-kasei.co.jp.
Shin-ich
SI
Sakasegawa
Department of Biochemistry, Teikyo University School of Medicine, 2-11-1 Kaga, Itabashi-Ku, Tokyo, 173-8605, Japan. maeba@med.teikyo-u.ac.jp.
Ryota
R
Maeba
Division of Biomedical Measurements and Diagnostics, Graduate School of Biomedical Engineering, Tohoku University, 2-1 Seiryo, Aoba, Sendai, 980-8575, Japan. kmura@bme.tohoku.ac.jp.
Kazutaka
K
Murayama
Asahi Kasei Pharma Corp., 632-1 Mifuku, Izunokuni, Shizuoka, 410-2321, Japan. matsumoto.hb@om.asahi-kasei.co.jp.
Hideyuki
H
Matsumoto
Department of Symbiotic Systems Science and Technology, Graduate School of Symbiotic Systems Science and Technology, Fukushima University, 1 Kanayagawa, Fukushima, 960-1296, Japan. sugimori@sss.fukushima-u.ac.jp.
Daisuke
D
Sugimori
D000818:Animals; D001161:Arteriosclerosis / Q000175:diagnosis; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D003704:Dementia / Q000175:diagnosis; D042241:Early Diagnosis; D006801:Humans; D006868:Hydrolysis; D008958:Models, Molecular; D054466:Phospholipases A1 / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010955:Plasmalogens / Q000737:chemistry*; D013302:Streptomyces / Q000201:enzymology*
D001426:Bacterial Proteins; D010955:Plasmalogens; D054466:Phospholipases A1
Arteriosclerosis; Dementia: diagnostic enzyme; Phospholipase A1; Plasmalogen; Plasmalogen determination; Streptomyces