Inhibition of hemostasis by a high affinity biogenic amine-binding protein from the saliva of a blood-feeding insect.
The saliva of the blood-feeding insect Rhodnius prolixus contains numerous pharmacologically active substances. Included among these are a number of lipocalin proteins that bind various ligands important in hemostasis and inflammation. One such protein is a biogenic amine-binding protein (ABP) that binds serotonin, epinephrine, and norepinephrine. Based on amino acid alignments, it is most similar to the nitrophorin group of lipocalins found in the same insect species. Physiologically, this protein appears to act as both a vasodilator and platelet aggregation inhibitor. This protein inhibits smooth muscle contraction of the rat uterus in response to serotonin and of the rabbit aorta in response to norepinephrine. Platelet aggregation induced by a combination of low concentrations of ADP and either serotonin or epinephrine is inhibited because of the binding of serotonin and epinephrine. Potentiation of aggregation induced by low concentrations of collagen along with serotonin or epinephrine is also inhibited. Dissociation constants for biogenic amines were measured using isothermal titration calorimetry and the Hummel-Dreyer method of equilibrium gel filtration. In this manner, K(d) values of 102, 24, and 345 nm were found for serotonin, norepinephrine, and epinephrine, respectively. Molecular modeling of ABP suggests that ligand binding is mediated by interaction with the side chains of aromatic amino acids and charged residues that line the binding pocket.
Andersen
John F
JF
Laboratory of Malaria and Vector Research, NIAID, National Institutes of Health, Bethesda, Maryland 20892, USA. janderson@niaid.nih.gov
Francischetti
Ivo M B
IM
Valenzuela
Jesus G
JG
Schuck
Peter
P
Ribeiro
Jose M C
JM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001679:Biogenic Amines / Q000378:metabolism; D004837:Epinephrine / Q000494:pharmacology; D006487:Hemostasis / Q000187:drug effects*; D006801:Humans; D019476:Insect Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009119:Muscle Contraction / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects; D009638:Norepinephrine / Q000494:pharmacology; D010974:Platelet Aggregation / Q000187:drug effects; D011485:Protein Binding; D051381:Rats; D012239:Rhodnius; D012471:Salivary Proteins and Peptides / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*
D001679:Biogenic Amines; D019476:Insect Proteins; D012471:Salivary Proteins and Peptides; D009638:Norepinephrine; D004837:Epinephrine