Presence in human eosinophils of a lysophospholipase similar to that found in the pancreas.
The supernatant fraction from lysed human eosinophils, when separated by gel-filtration chromatography, contains a protein with lysophospholipase activity of approximate molecular mass 74 kDa. This mass differs substantially from the 17 kDa of a previously cloned eosinophil lysophospholipase (Charcot-Leyden crystal protein), but is similar to that reported for a pancreatic enzyme. We have therefore further characterized this pancreatic-like lysophospholipase in human eosinophils. A rabbit polyclonal antibody was produced against a synthetic peptide consisting of amino acids 325-349 from the 74 kDa rat pancreatic lysophospholipase. Western-blot analysis of eosinophil extracts indicate that this antibody recognizes a single 74 kDa band in these preparations. Incubation of the supernatant fraction from sonified eosinophils with this antibody, followed by precipitation of antibody-antigen complexes with Protein A, removes the majority of the lysophospholipase activity. Indirect immunofluorescence examination with this antibody indicates this protein to be localized to granules of eosinophils and not in other leucocytes. Moreover, reverse transcriptase PCR of polyadenylated RNA from eosinophils and from rat pancreatic tissue with primers to rat pancreatic lysophospholipase resulted in readily detectable 1 kb DNA products in both samples. Sequencing revealed this DNA fragment to be identical with the human pancreatic lysophospholipase cDNA sequence. Taken together, these data indicate that eosinophils contain a lysophospholipase that is similar to the human pancreatic enzyme.
Holtsberg
F W
FW
Schering Plough Research Institute, Kenilworth, NJ 07033, USA.
Ozgur
L E
LE
Garsetti
D E
DE
Myers
J
J
Egan
R W
RW
Clark
M A
MA
D000595:Amino Acid Sequence; D000818:Animals; D004804:Eosinophils / Q000201:enzymology*; D006801:Humans; D008245:Lysophospholipase / Q000032:analysis*; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D051381:Rats
D008245:Lysophospholipase