Characterization of recombinant Streptococcus mitis-derived human platelet aggregation factor

APMIS. 2012 Jan;120(1):56-71. doi: 10.1111/j.1600-0463.2011.02813.x. Epub 2011 Sep 27.

Abstract

We previously purified Streptococcus mitis-derived human platelet aggregation factor (Sm-hPAF) from the culture supernatant of S. mitis strain Nm-65, isolated from the tooth surface of a patient with Kawasaki disease. Here we produced recombinant Sm-hPAF protein (rSm-hPAF) in Escherichia coli, to determine whether rSm-hPAF conserves its platelet aggregation activity. rSm-hPAF precursor (665 amino acids) shows up to 36-56% identity with the family of cholesterol-dependent cytolysins (CDCs), and rSm-hPAF displayed potent hemolytic activity toward mammalian erythrocytes, including human erythrocytes with platelet aggregation activity. The 162-amino acid amino-terminal domain of rSm-hPAF was found in no other CDCs except lectinolysin; this domain is homologous to a portion of pneumococcal fucolectin-related protein. Interestingly, suilysin (SLY) and pneumolysin (PLY) of CDCs also exhibit substantial human platelet aggregation activity, similar to rSm-hPAF, and the platelet aggregation by rSm-hPAF, SLY, and PLY was morphologically confirmed using light and electron microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal
  • Bacterial Proteins / toxicity
  • Bacterial Toxins / chemistry*
  • Bacterial Toxins / genetics
  • Bacterial Toxins / toxicity*
  • Base Sequence
  • Blood Platelets / drug effects
  • Blood Platelets / metabolism
  • Blood Platelets / ultrastructure
  • Cell Adhesion Molecules / chemistry*
  • Cell Adhesion Molecules / genetics
  • Cell Adhesion Molecules / toxicity*
  • Cholesterol / chemistry
  • Cytotoxins / chemistry*
  • Cytotoxins / genetics
  • Cytotoxins / toxicity*
  • DNA, Bacterial / genetics
  • Female
  • Genes, Bacterial
  • Hemolysin Proteins / toxicity
  • Humans
  • In Vitro Techniques
  • L-Lactate Dehydrogenase / blood
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Electron, Transmission
  • Molecular Sequence Data
  • Mucocutaneous Lymph Node Syndrome / blood
  • Mucocutaneous Lymph Node Syndrome / etiology
  • Mucocutaneous Lymph Node Syndrome / microbiology
  • Phylogeny
  • Platelet Aggregation / drug effects*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / toxicity
  • Sequence Homology, Amino Acid
  • Streptococcus mitis / genetics
  • Streptococcus mitis / pathogenicity*
  • Streptococcus mitis / physiology*
  • Streptolysins / toxicity

Substances

  • Antibodies, Monoclonal
  • Bacterial Proteins
  • Bacterial Toxins
  • Cell Adhesion Molecules
  • Cytotoxins
  • DNA, Bacterial
  • Hemolysin Proteins
  • Recombinant Proteins
  • Streptolysins
  • cell aggregation factors
  • plY protein, Streptococcus pneumoniae
  • suilysin
  • Cholesterol
  • L-Lactate Dehydrogenase