Abstract
PL37 (RAARISLGPRCIKAFTE) is an antisense homology box peptide composed of aa 37-53 of C5a-anaphylatoxin and is considered to be the region essential for C5a function. Using a computer program, we designed the complementary peptides ASGAPAPGPAGPLRPMF (Pep-A) and ASTAPARAGLPRLPKFF (Pep-B). Pep-A bound to PL37 and to C5a with very slow dissociation as determined by analysis using surface plasmon resonance, whereas Pep-B failed to bind at all. C5a was inactivated by concentrations of 7 nM or more of Pep-A, and this concentration of Pep-A inhibited induction of intracellular Ca(2+) influx in neutrophils. Patch clamp electrophysiology experiments also showed the effectiveness of Pep-A in C5aR-expressing neuroblastoma cells. Furthermore, Pep-A administration prevented rats from C5a-mediated rapid lethal shock induced by an Ab to a membrane inhibitor of complement after LPS sensitization.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Anaphylaxis / mortality
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Anaphylaxis / prevention & control
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Animals
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Antigens / chemistry
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Antigens / metabolism
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Antigens / physiology
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Antisense Elements (Genetics) / chemical synthesis
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Antisense Elements (Genetics) / metabolism
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Antisense Elements (Genetics) / physiology
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Calcium / metabolism
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Cell Line, Tumor
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Cell Separation
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Complement C5a / antagonists & inhibitors*
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Complement C5a / metabolism*
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Complement C5a / physiology
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Complement Inactivator Proteins / chemical synthesis
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Complement Inactivator Proteins / metabolism
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Complement Inactivator Proteins / physiology*
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Disease Models, Animal
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Humans
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Intracellular Fluid / metabolism
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Male
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Molecular Sequence Data
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Neutrophils / immunology
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Neutrophils / metabolism
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Patch-Clamp Techniques
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Peptide Fragments / chemical synthesis
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Peptide Fragments / metabolism
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Peptide Fragments / physiology*
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Protein Binding / immunology
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Rats
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Rats, Wistar
Substances
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Antigens
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Antisense Elements (Genetics)
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Complement Inactivator Proteins
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Peptide Fragments
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Complement C5a
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Calcium