Recognition of anionic phospholipid membranes by an antihemostatic protein from a blood-feeding insect.
The saliva of blood-feeding insects contains a variety of molecules having antihemostatic activity. Here, we describe nitrophorin 7 (NP7), a salivary protein that binds with high affinity to anionic phospholipid membranes. The protein is apparently targeted to the negatively charged surfaces of activated platelets and other cells, where it can serve as a vasodilator, antihistamine, platelet aggregation inhibitor, and anticoagulant. As with other members of the nitrophorin group, NP7 reversibly binds a molecule of NO and binds histamine with high affinity. The protein differs from other nitrophorins in that it binds to membranes containing phosphatidylserine. Sedimentation and surface plasmon resonance experiments, revealed two classes of phospholipid-binding sites having K(d) values of 4.8 and 755 nM. NP7 inhibits prothrombin activation by blocking phospholipid binding sites for the prothrombinase complex on the surfaces of vesicles and activated platelets. As a NO complex, NP7 inhibits collagen and ADP-induced platelet aggregation and induces disaggregation of ADP-stimulated platelets by an NO-mediated mechanism. Molecular modeling of NP7 revealed a putative, positively charged membrane interaction surface comprised mainly of a helix lying outside of the lipocalin beta-barrel structure.
Andersen
John F
JF
Laboratory of Malaria and Vector Research, National Institutes of Health, NIAID, Rockville, Maryland 20852, USA. jandersen@niaid.nih.gov
Gudderra
Nanda P
NP
Francischetti
Ivo M B
IM
Valenzuela
Jesus G
JG
Ribeiro
José M C
JM
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001777:Blood Coagulation; D001792:Blood Platelets / Q000378:metabolism*; D002462:Cell Membrane / Q000502:physiology*; D006420:Hemeproteins / Q000378:metabolism / Q000494:pharmacology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010718:Phosphatidylserines / Q000737:chemistry / Q000378:metabolism*; D010974:Platelet Aggregation / Q000187:drug effects; D011516:Prothrombin / Q000037:antagonists & inhibitors*; D012469:Salivary Glands / Q000737:chemistry*; D012471:Salivary Proteins and Peptides / Q000378:metabolism / Q000494:pharmacology*; D017386:Sequence Homology, Amino Acid; D012965:Sodium Chloride / Q000737:chemistry / Q000378:metabolism; D020349:Surface Plasmon Resonance; D013925:Thromboplastin / Q000378:metabolism / Q000494:pharmacology
D006420:Hemeproteins; D010718:Phosphatidylserines; D012471:Salivary Proteins and Peptides; C090560:nitrophorin; D012965:Sodium Chloride; D011516:Prothrombin; D013925:Thromboplastin