Melittin binds to secretory phospholipase A2 and inhibits its enzymatic activity

Shamsher S. Saini, Johnny Peterson, Ashok Chopra

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Synthetic melittin inhibited the enzymatic activity of secretory phospholipase A2 (PLA2) from various sources, including bee and snake venoms, bovine pancreas, and synovial fluid from rheumatoid arthritis patients, irrespective of substrate (e,g., [14C]-phosphatidylcholine or phosphatidylethanolamine vesicles and [3H]-oleic acid-labeled E.coli). A Lineweaver-Burk analysis showed that melittin was a noncompetitive inhibitor of bee venom PLA2, causing a change in V(max) from 200 to 50 units/min/mg of protein. The K(m) remained unchanged (0.75 nmole). Melittin inhibited approximately 50% of purified bee venom PLA2 activity in a 30:1 molar ratio (melittin:enzyme). Because the enzyme kinetics indicated a PLA2-melittin interaction, a melittin-sepharose affinity column was used to purify a PLA2 from human serum. Further, an enzyme-linked assay was developed to quantitate PLA2 activity in biological fluids using avidin-peroxidase and ELISA plates coated with biotinylated melittin. These observations may have potential therapeutic significance, as well as provide a convenient basis for the isolation and quantitation of PLA2.

Original languageEnglish (US)
Pages (from-to)436-442
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume238
Issue number2
DOIs
StatePublished - Sep 18 1997

Fingerprint

Secretory Phospholipase A2
Melitten
Phospholipases A2
Bee Venoms
Enzyme kinetics
Snake Venoms
Fluids
Avidin
Synovial Fluid
Enzyme Assays
Enzymes
Oleic Acid
Phosphatidylcholines
Sepharose
Escherichia coli
Peroxidase
Pancreas
Assays
Rheumatoid Arthritis
Enzyme-Linked Immunosorbent Assay

Keywords

  • Bee venom
  • Bovine pancreas
  • Enzyme kinetics
  • Fatty acid
  • Inflammation
  • Melittin
  • Phospholipase A
  • Phospholipid
  • Rheumatoid arthritis
  • Snake venom
  • Synovial fluid

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Melittin binds to secretory phospholipase A2 and inhibits its enzymatic activity. / Saini, Shamsher S.; Peterson, Johnny; Chopra, Ashok.

In: Biochemical and Biophysical Research Communications, Vol. 238, No. 2, 18.09.1997, p. 436-442.

Research output: Contribution to journalArticle

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