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KMID : 0364519960080020179
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Dong-A Journal Medicine
1996 Volume.8 No. 2 p.179 ~ p.188
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Effects of Antibody against Consensus Sequence of Carboxyl Terminal Region from Phospholipase d on the Enzyme from Human Neutrophil
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Abstract
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guanosine 5'-O-(3-thiotriphosphate) (GTPγS)-dependent phospholipase D activity in human neutrophils requires protein factors in both plasma membrane and cytosol. The activity of cytosolic phospholipase d was investigated using exogenous
phospholipid as
a substrate. Phospholipase D has the most conserved sequence in the carboxyl-terminal region of the enzyme. The consensus sequence is also found in P37 from vaccinia virus which protein is known to have an important function role in the viral
infectivity, We used antibody against the peptide carboxyl-terminal consensus region to know which protein is phospholipase D from human neutrophil. The antibody could detect an 100 kD protein from a cytosolic fraction, not from a plasma
memnbrane
and
immunoprecipitate the cytosolic protein specifically. The protein was detected in flow-through fraction using DEAE-Cellulose column and separated from the cytosolic phsopholipase D activity. It did not have inhibitory effect on the activation of
plasma
membrane-and cytosol-associated phospholipase D and cytosolic phospholipase D. Immunoprecipitate from the cytosol of neutrophil using the antibody had no phospholipase D activity and immunodepleted cytosol showed the activity.
These data suggest that an 100kD protein is not phospholipase D in human neutrophil and the consensus sequence is not specific to phospholipase. D.
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KEYWORD
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