PMID- 10085103
OWN - NLM
STAT- MEDLINE
DCOM- 19990429
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 13
DP  - 1999 Mar 26
TI  - Purification and characterization from rat kidney membranes of a novel
      platelet-activating factor (PAF)-dependent transacetylase that catalyzes the
      hydrolysis of PAF, formation of PAF analogs, and C2-ceramide.
PG  - 8655-61
AB  - We have previously identified two enzyme activities that transfer the acetyl
      group from platelet-activating factor (PAF) in a CoA-independent manner to
      lysoplasmalogen or sphingosine in HL-60 cells, endothelial cells, and a variety
      of rat tissues. These were termed as PAF:lysoplasmalogen (lysophospholipid)
      transacetylase and PAF:sphingosine transacetylase, respectively. In the present
      study, we have solubilized and purified this PAF-dependent transacetylase
      13,700-fold from rat kidney membranes (mitochondrial plus microsomal membranes)
      based on the PAF:lysoplasmalogen transacetylase activity. The mitochondria and
      microsomes were prepared and washed three times, then solubilized with 0.04%
      Tween 20 at a detergent/protein (w/w) ratio of 0.1. The solubilized fractions
      from mitochondria and microsomes were combined and subjected to sequential column
      chromatographies on DEAE-Sepharose, hydroxyapatite, phenyl-Sepharose, and
      chromatofocusing. The enzyme was further purified by native-polyacrylamide gel
      electrophoresis (PAGE) and affinity gel matrix in which the competitive inhibitor
      of the enzyme, 1-O-hexadecyl-2-N-methylcarbamyl-sn-glycero-3-phosphoethanolamine 
      was covalently attached to the CH-Sepharose. On SDS-PAGE, the purified enzyme
      showed a single homogeneous band with an apparent molecular mass of 40 kDa. The
      purified enzyme catalyzed transacetylation of the acetyl group not only from PAF 
      to lysoplasmalogen forming plasmalogen analogs of PAF, but also to sphingosine
      producing N-acetylsphingosine (C2-ceramide). In addition, this enzyme acted as a 
      PAF-acetylhydrolase in the absence of lipid acceptor molecules. These results
      suggest that PAF-dependent transacetylase is an enzyme that modifies the cellular
      functions of PAF through generation of other diverse lipid mediators.
FAU - Karasawa, K
AU  - Karasawa K
AD  - Biochemistry, Basic and Applied Research Unit, Oak Ridge Associated Universities,
      Oak Ridge, Tennessee 37831-0117, USA.
FAU - Qiu, X
AU  - Qiu X
FAU - Lee, T
AU  - Lee T
LA  - eng
GR  - HL-52492/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Ceramides)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Ethanolamines)
RN  - 0 (Plasmalogens)
RN  - 0 (Platelet Activating Factor)
RN  - 0 (Polysorbates)
RN  - EC 2.3.1.- (Acetyltransferases)
RN  - EC 2.3.1.- (sphingosine CoA-independent transacetylase)
RN  - NGZ37HRE42 (Sphingosine)
SB  - IM
MH  - Acetyltransferases/*chemistry
MH  - Animals
MH  - Cell Membrane/enzymology
MH  - Ceramides/*metabolism
MH  - Chromatography, Affinity
MH  - Enzyme Inhibitors/pharmacology
MH  - Ethanolamines/pharmacology
MH  - Kidney/*enzymology
MH  - Kinetics
MH  - Microsomes/enzymology
MH  - Mitochondria/enzymology
MH  - Plasmalogens/metabolism
MH  - Platelet Activating Factor/*metabolism
MH  - Polysorbates
MH  - Rats
MH  - Sphingosine/metabolism
MH  - Substrate Specificity
EDAT- 1999/03/20 00:00
MHDA- 1999/03/20 00:01
CRDT- 1999/03/20 00:00
PHST- 1999/03/20 00:00 [pubmed]
PHST- 1999/03/20 00:01 [medline]
PHST- 1999/03/20 00:00 [entrez]
AID - 10.1074/jbc.274.13.8655 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Mar 26;274(13):8655-61. doi: 10.1074/jbc.274.13.8655.