Purification and characterization of prolixin S (nitrophorin 2), the salivary anticoagulant of the blood-sucking bug Rhodnius prolixus.
The salivary anticoagulant of the blood-sucking bug Rhodnius prolixus was purified to homogeneity using a protocol consisting of weak cation-exchange, DEAE, hydrophobic-interaction and octadecyl reverse-phase chromatography, yielding a protein with the same N-terminal sequence as nitrophorin 2, one of the four NO haem protein carriers present in the salivary glands of Rhodnius with a molecular mass of 19689 Da [D. Champagne, R.H. Nussenzveig and J.M.C. Ribeiro, (1995) J. Biol. Chem. 270, in the press]. To exclude the possibility of the nitrophorin being a contaminant, another chromatographic protocol was performed, consisting of chromatofocusing followed by strong-cation-exchange chromatography. Again the anticoagulant was eluted with nitrophorin 2. Nitrophorin 2 inhibits coagulation Factor VIII-mediated activation of Factor X and accounts for all the anti-clotting activity observed in Rhodnius salivary glands.
Ribeiro
J M
JM
Department of Entomology, University of Arizona, Tucson 85721, USA.
Schneider
M
M
Guimarães
J A
JA
D000595:Amino Acid Sequence; D000818:Animals; D000925:Anticoagulants / Q000302:isolation & purification*; D001777:Blood Coagulation; D015944:Factor VIIIa / Q000378:metabolism; D005170:Factor X / Q000378:metabolism; D006420:Hemeproteins / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D012239:Rhodnius / Q000737:chemistry*; D012471:Salivary Proteins and Peptides / Q000737:chemistry / Q000302:isolation & purification*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013917:Thrombin / Q000378:metabolism
D000925:Anticoagulants; D006420:Hemeproteins; D012471:Salivary Proteins and Peptides; C090560:nitrophorin; D015944:Factor VIIIa; D005170:Factor X; D013917:Thrombin