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.
© 2011 The Authors. APMIS © 2011 APMIS.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adult
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal
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Bacterial Proteins / toxicity
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Bacterial Toxins / chemistry*
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Bacterial Toxins / genetics
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Bacterial Toxins / toxicity*
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Base Sequence
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Blood Platelets / drug effects
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Blood Platelets / metabolism
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Blood Platelets / ultrastructure
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Cell Adhesion Molecules / chemistry*
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Cell Adhesion Molecules / genetics
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Cell Adhesion Molecules / toxicity*
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Cholesterol / chemistry
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Cytotoxins / chemistry*
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Cytotoxins / genetics
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Cytotoxins / toxicity*
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DNA, Bacterial / genetics
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Female
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Genes, Bacterial
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Hemolysin Proteins / toxicity
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Humans
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In Vitro Techniques
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L-Lactate Dehydrogenase / blood
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Mice
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Mice, Inbred BALB C
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Microscopy, Electron, Transmission
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Molecular Sequence Data
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Mucocutaneous Lymph Node Syndrome / blood
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Mucocutaneous Lymph Node Syndrome / etiology
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Mucocutaneous Lymph Node Syndrome / microbiology
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Phylogeny
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Platelet Aggregation / drug effects*
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / toxicity
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Sequence Homology, Amino Acid
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Streptococcus mitis / genetics
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Streptococcus mitis / pathogenicity*
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Streptococcus mitis / physiology*
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Streptolysins / toxicity
Substances
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Antibodies, Monoclonal
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Bacterial Proteins
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Bacterial Toxins
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Cell Adhesion Molecules
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Cytotoxins
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DNA, Bacterial
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Hemolysin Proteins
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Recombinant Proteins
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Streptolysins
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cell aggregation factors
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plY protein, Streptococcus pneumoniae
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suilysin
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Cholesterol
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L-Lactate Dehydrogenase