The hydrolysis of the alkenyl group from enthanolamine-plasmalogen by oligodendroglial cell-enriched fractions

Freeman, Neil M.; Carey, Eric M.; Carruthers, Angela

doi:10.1016/0197-0186(84)90103-7

Cited by 4 publications

(2 citation statements)

References 30 publications

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“…These workers found no evidence for an enzyme that catalyzed hydrolysis of the vinyl ether group of the intact plasmalogen molecule in any subcellular fraction of rat brain (25). A lysoplasmalogenase activity was also identified in oligodendroglial cell enriched fractions from bovine brain that had properties similar to those of purified liver lysoplasmalogenase in its neutral pH optimum and lack of requirement for Mg 2+ (47). A lysoplasmalogenase activity was reported in guinea pig heart microsomes and mitochondria, using a coupled enzyme assay with aldehyde dehydrogenase as coupling enzyme (48).…”

Section: Discussionmentioning

confidence: 94%

Purification, Identification, and Cloning of Lysoplasmalogenase, the Enzyme That Catalyzes Hydrolysis of the Vinyl Ether Bond of Lysoplasmalogen

Pfeiffer

Calhoon

et al. 2011

Journal of Biological Chemistry

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Lysoplasmalogenase (EC 3.3.2.2 and EC 3.3.2.5) is an enzyme that catalyzes hydrolytic cleavage of the vinyl ether bond of lysoplasmalogen, forming fatty aldehyde and glycerophosphoethanolamine or glycerophosphocholine and is specific for the sn-2-deacylated form of plasmalogen. Here we report the purification, characterization, identification, and cloning of lysoplasmalogenase. Rat liver microsomal lysoplasmalogenase was solubilized with octyl glucoside and purified 500-fold to near homogeneity using four chromatography steps. The purified enzyme has apparent Km values of ∼50 μm for both lysoplasmenylcholine and lysoplasmenylethanolamine and apparent Vm values of 24.5 and 17.5 μmol/min/mg protein for the two substrates, respectively. The pH optimum was 7.0. Lysoplasmalogenase was competitively inhibited by lysophosphatidic acid (Ki ∼20 μm). The predominant band on a gel at ∼19 kDa was subjected to trypsinolysis, and the peptides were identified by mass spectrometry as Tmem86b, a protein of unknown function. Transient transfection of human embryonic kidney (HEK) 293T cells showed that TMEM86b cDNA yielded lysoplasmalogenase activity, and Western blot analyses confirmed the synthesis of TMEM86b protein. The protein was localized in the membrane fractions. The TMEM86b gene was also transformed into Escherichia coli, and its expression was verified by Western blot and activity analyses. Tmem86b is a hydrophobic transmembrane protein of the YhhN family. Northern blot analyses demonstrated that liver expressed the highest level of Tmem86b, which agreed with tissue distribution of activity. Overexpression of TMEM86b in HEK 293T cells resulted in decreased levels of plasmalogens, suggesting that the enzyme may be important in regulating plasmalogen levels in animal cells.

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Section: Discussionmentioning

confidence: 94%

Purification, Identification, and Cloning of Lysoplasmalogenase, the Enzyme That Catalyzes Hydrolysis of the Vinyl Ether Bond of Lysoplasmalogen

Pfeiffer

Calhoon

et al. 2011

Journal of Biological Chemistry

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“…Type II enzymes, of which isochorismatase is an example, function as hydrolases to cleave a vinyl ether bond, releasing both alcohol and carbonyl products (Scheme III). Besides isochorismatase, the only other example we are aware of at this time is the enzyme plasmalogenase, present in liver microsomes (Warner & Lands, 1961; Ellingson & Lands, 1968; Gunawan & Debuch, 1981) and oligodendroglial brain cells (Freeman et al, 1984). Plasmalogenase hydrolyzes the alkenyl ether bond of l-0-(l,-alkenyl)-2-0-acyl-5n-glycero-3phosphoethanolamine to produce a long-chain aldehyde and the 2-acylglycerophosphoethanolamine.…”

Section: Discussionmentioning

confidence: 99%

Subcloning of the enterobactin biosynthetic gene entB: expression, purification, characterization and substrate specificity of isochorismatase

Rusnak¹,

Liu²,

Quinn³

et al. 1990

Biochemistry

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The Escherichia coli entB gene, coding for the enterobactin biosynthetic enzyme isochorismatase, has been subcloned into the multicopy plasmid pKK223-3 under the control of the tac promoter. The resulting recombinant plasmid pFR1 expresses isochorismatase amounting to over 50% of the total cellular protein. The enzyme has been purified to homogeneity and a convenient assay developed. The enzyme has a Km for isochorismate of 14.7 microM and a turnover number of 600 min-1. By use of 1H NMR spectroscopy, the progress of the reaction was followed with the expected formation of 2,3-dihydro-2,3-dihydroxybenzoate product. Several substrate analogues were also utilized by the enzyme including chorismic acid, the immediate precursor to isochorismic acid in the enterobactin biosynthetic pathway.

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Biosynthetic preparation of radioactively labelled ethanolamine plasmalogen (1-O-[1′-14C]octadec-1′-enyl-2-acyl-sn-glycero-3-phosphoethanolamine) using a protozoan cell culture

Achterberg¹,

Fricke²,

Gercken³

1986

Chemistry and Physics of Lipids

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The hydrolysis of the alkenyl group from enthanolamine-plasmalogen by oligodendroglial cell-enriched fractions

Cited by 4 publications

References 30 publications

Purification, Identification, and Cloning of Lysoplasmalogenase, the Enzyme That Catalyzes Hydrolysis of the Vinyl Ether Bond of Lysoplasmalogen

Purification, Identification, and Cloning of Lysoplasmalogenase, the Enzyme That Catalyzes Hydrolysis of the Vinyl Ether Bond of Lysoplasmalogen

Subcloning of the enterobactin biosynthetic gene entB: expression, purification, characterization and substrate specificity of isochorismatase

Biosynthetic preparation of radioactively labelled ethanolamine plasmalogen (1-O-[1′-14C]octadec-1′-enyl-2-acyl-sn-glycero-3-phosphoethanolamine) using a protozoan cell culture

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