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. 2012 Jun 13;17(3):459–478. doi: 10.2478/s11658-012-0019-2

Differences between group X and group V secretory phospholipase A2 in lipolytic modification of lipoproteins

Shigeki Kamitani 1,2,✉, Katsutoshi Yamada 2, Shigenori Yamamoto 2, Yoshikazu Ishimoto 2, Takashi Ono 2, Akihiko Saiga 2, Kohji Hanasaki 2
PMCID: PMC6275602  PMID: 22706677

Abstract

Secretory phospholipases A2 (sPLA2s) are a diverse family of low molecular mass enzymes (13–18 kDa) that hydrolyze the sn-2 fatty acid ester bond of glycerophospholipids to produce free fatty acids and lysophospholipids. We have previously shown that group X sPLA2 (sPLA2-X) had a strong hydrolyzing activity toward phosphatidylcholine in low-density lipoprotein (LDL) linked to the formation of lipid droplets in the cytoplasm of macrophages. Here, we show that group V sPLA2 (sPLA2-V) can also cause the lipolysis of LDL, but its action differs remarkably from that of sPLA2-X in several respects. Although sPLA2-V released almost the same amount of fatty acids from LDL, it released more linoleic acid and less arachidonic acid than sPLA2-X. In addition, the requirement of Ca2+ for the lipolysis of LDL was about 10-fold higher for sPLA2-V than sPLA2-X. In fact, the release of fatty acids from human serum was hardly detectable upon incubation with sPLA2-V in the presence of sodium citrate, which contrasted with the potent response to sPLA2-X. Moreover, sPLA2-X, but not sPLA2-V, was found to specifically interact with LDL among the serum proteins, as assessed by gel-filtration chromatography as well as sandwich enzyme-immunosorbent assay using anti-sPLA2-X and anti-apoB antibodies. Surface plasmon resonance studies have revealed that sPLA2-X can bind to LDL with high-affinity (Kd = 3.1 nM) in the presence of Ca2+. Selective interaction of sPLA2-X with LDL might be involved in the efficient hydrolysis of cell surface or intracellular phospholipids during foam cell formation.

Key words: Secretory phospholipase A2, Low-density lipoprotein, High-density lipoprotein, Phospholipids, Calcium ion

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Abbreviations used

Ab

antibody

apoB

apolipoprotein B

BSA

bovine serum albumin

COX

cyclooxygenase

FCS

fetal calf serum

HDL

high density lipoprotein

HPLC

high-performance liquid chromatography

LDL

low density lipoprotein

lysoPC

lysophosphatidylcholine

PBS

phosphate-buffered saline

PC

phosphatidylcholine

PLA2

phospholipase A2

SDS-PAGE

SDS-polyacrylamide gel electrophoresis

sPLA2

secretory PLA2

sPLA2-IB

group IB sPLA2

sPLA2-IIA

group IIA sPLA2

sPLA2-V

group V sPLA2

sPLA2-X

group X sPLA2

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