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Characterization of two distinct phospholipase C enzymes from Burkholderia pseudomallei

Sunee Korbsrisate1, Andrew P Tomaras2, Suwat Damnin1

  • 1Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.

Microbiology (Reading, England)
|May 29, 2007
PubMed

Insights

Two phospholipase C (PLC) enzymes, Plc-1 and Plc-2, from Burkholderia pseudomallei contribute to virulence. Characterizing these enzymes reveals they are not redundant and play distinct roles in host cell interactions during melioidosis.

Area of Science:

  • Microbiology
  • Bacterial Pathogenesis
  • Enzymology

Background:

  • Burkholderia pseudomallei causes melioidosis, a severe human infection.
  • Phospholipase C (PLC) enzymes are potential virulence factors in bacterial pathogens.

Purpose of the Study:

  • To characterize the roles of two specific PLC enzymes (Plc-1 and Plc-2) from B. pseudomallei in virulence.
  • To determine if Plc-1 and Plc-2 are redundant or have distinct functions.

Main Methods:

  • Construction and analysis of B. pseudomallei single and double plc mutants.
  • Assays for extracellular PLC activity, phospholipid utilization, and hemolytic activity.
  • Infection of eukaryotic cells to assess plaque formation, cytotoxicity, multinucleated giant cell (MNGC) formation, and apoptosis induction.

Main Results:

  • Single plc mutants showed reduced extracellular PLC activity, confirming functional extracellular PLCs.
  • Both Plc-1 and Plc-2 contributed to phospholipid utilization but were not hemolytic.
  • Plc-1 enhanced Plc-2's plaque-forming efficiency; Plc-2 significantly contributed to cytotoxicity.
  • Neither enzyme was involved in MNGC formation or apoptosis induction.

Conclusions:

  • B. pseudomallei Plc-1 and Plc-2 are non-redundant virulence factors.
  • These PLCs contribute to B. pseudomallei pathogenesis through distinct mechanisms, including phospholipid hydrolysis and host cell cytotoxicity.

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