The 85-kDa, arachidonic acid-specific phospholipase A2 is expressed as an activated phosphoprotein in Sf9 cells.

Abstract:

:A human, 85-kDa arachidonoyl-specific, cytosolic phospholipase A2 was expressed using the baculovirus-insect cell expression system. Expression resulted in the production of an active protein which consisted of approximately 3% of the total protein in the host Spodoptera frugiperda (Sf9) cells at 67 h after infection. The phospholipase A2 was purified to apparent homogeneity and exhibited calcium-dependent phospholipase A2 activity with a specific activity of 8 mumol/min/mg protein, as well as calcium-independent lysophospholipase activity with a specific activity of 17 mumol/min/mg protein. The phospholipase A2 was expressed as a phosphoprotein and was primarily phosphorylated on serine residues. Phosphatase treatment of the recombinant phospholipase A2 resulted in dephosphorylation of the enzyme and a 63% decrease in phospholipase A2 activity. This decrease in activity is similar in magnitude to the decrease in activity observed with phosphatase-treated phospholipase A2 from stimulated mammalian cells. These data demonstrate that the 85-kDa phospholipase A2 is expressed as an activated phosphoprotein in Sf9 cells.

journal_name

Arch Biochem Biophys

authors

de Carvalho MS,McCormack FX,Leslie CC

doi

10.1006/abbi.1993.1549

subject

Has Abstract

pub_date

1993-11-01 00:00:00

pages

534-40

issue

2

eissn

0003-9861

issn

1096-0384

pii

S0003986183715493

journal_volume

306

pub_type

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