Melittin-mediated release of [3H]-oleic acid from E. coli cells is dependent upon heat- and trypsin-sensitive factor(s) in human serum

Shamsher S. Saini, Ashok Chopra, Johnny Peterson

Research output: Contribution to journalArticle

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Abstract

Synthetic melittin mediated the release of [3H]-oleic acid ([3H]-OA) or its acylated lipids from [3H]-OA-labeled E. coli cells exposed to human serum. This phenomenon was not observed in the absence of serum and was calcium independent. The addition of serum was not required for melittin-mediated lysis of erythrocytes, although lysis was greater in the presence of serum than in its absence (P<0.001). Trypsin treatment of human serum reduced the melittin-mediated release of [3H]-OA/acylated lipids, and this effect was more pronounced upon boiling the serum (P<0.01). A kinetic study showed that maximum release of [3H]-OA/acylated lipids occurred within 3-6 min. Thin layer chromatography (TLC) analysis showed the lipids to be phosphatidyl ethanolamine (PE), phosphatidylethanol (PEt) and phosphatidic acid (PA). There was no detectable level of oleic acid (OA), diacylglycerol (DAG), phosphatidyl choline (PC) or phosphatidyl serine (PS). These findings suggested that a trypsin and heat-sensitive enzyme/factor present in the serum had a role in melittin-mediated action. These findings further showed that melittin activated phospholipase D (PLD), without affecting phospholipase A2 (PLA2) or phospholipase C (PLC) activity. Copyright (C) 2000 Elsevier Science Ltd.

Original languageEnglish (US)
Pages (from-to)1077-1086
Number of pages10
JournalToxicon
Volume38
Issue number8
DOIs
StatePublished - Aug 1 2000

Fingerprint

Melitten
Oleic Acid
Trypsin
Escherichia coli
Hot Temperature
Lipids
Serum
Thin layer chromatography
Phospholipase D
Phosphatidic Acids
Ethanolamine
Phospholipases A2
Phosphatidylserines
Diglycerides
Type C Phospholipases
Phosphatidylcholines
Boiling liquids
Thin Layer Chromatography
Calcium
Kinetics

Keywords

  • Human serum
  • Melittin
  • Phospholipase A, C and D
  • Phospholipids

ASJC Scopus subject areas

  • Toxicology

Cite this

Melittin-mediated release of [3H]-oleic acid from E. coli cells is dependent upon heat- and trypsin-sensitive factor(s) in human serum. / Saini, Shamsher S.; Chopra, Ashok; Peterson, Johnny.

In: Toxicon, Vol. 38, No. 8, 01.08.2000, p. 1077-1086.

Research output: Contribution to journalArticle

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