Characterization of immunoaffinity purified peptidoglycan-associated lipoprotein of Actinobacillus

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Isolation and Chemical Characterization of Lipid A from Gram-negative Bacteria
12:57

Isolation and Chemical Characterization of Lipid A from Gram-negative Bacteria

Published on: September 16, 2013

Riikka Ihalin1, Maribasappa Karched, Kjell Eneslätt

  • 1Division of Oral Microbiology, Umeå University, Umeå SE-90185, Sweden.

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
|December 28, 2005
PubMed

Insights

Researchers purified peptidoglycan-associated lipoprotein (PAL) from the periodontal pathogen Actinobacillus actinomycetemcomitans. This protein was recognized by patient sera, suggesting its role in infection and inflammation.

Area of Science:

  • Microbiology
  • Immunology
  • Bacterial Pathogenesis

Background:

  • Peptidoglycan-associated lipoprotein (PAL) is a conserved outer membrane protein in Gram-negative bacteria.
  • PAL can be proinflammatory and is sometimes released extracellularly.

Purpose of the Study:

  • To purify a novel PAL (AaPAL) from Actinobacillus actinomycetemcomitans.
  • To assess the immunogenicity of AaPAL in patients with A. actinomycetemcomitans infections.

Main Methods:

  • Immunoaffinity chromatography using AaPAL-specific antipeptide antibodies was employed for purification.
  • Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting were used to confirm purity and recognition.

Main Results:

  • A highly purified AaPAL was obtained, free from contaminating A. actinomycetemcomitans proteins.
  • Sera from patients infected with A. actinomycetemcomitans demonstrated recognition of the purified AaPAL.

Conclusions:

  • The purification method is effective for AaPAL and potentially other bacterial PALs.
  • AaPAL is immunogenic in infected individuals and may play a role in the proinflammatory mechanisms of A. actinomycetemcomitans.

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