Purification, characterization and cDNA cloning of a novel anticoagulant of the intrinsic pathway, (prolixin-S) from salivary glands of the blood sucking bug, Rhodnius prolixus.
The salivary glands of the blood sucking insect, Rhodnius prolixus, have an anticoagulant, prolixin-S, which was reported as a specific inhibitor of intrinsic coagulant pathway. Prolixin-S was purified from the salivary glands extract of Rhodnius prolixus by gel filtration and anion exchange HPLC by assaying prolongation of activated partial thromboplastin time (APTT). The isolated protein specifically inhibited factor IXa-catalyzed activation of factor X in the presence of Ca2+ and phospholipids irrespective of the presence or absence of factor VIIIa. The anticoagulant factor had red color and a specific absorbance peak at 402 nm and thus it was identified as a heme protein. A Rhodnius prolixus salivary gland cDNA library was prepared, screened with an antibody against prolixin-S and its complete cDNA sequence was determined. cDNA and deduced amino acid sequences showed that prolixin-S is a novel anticoagulant of 19,922 Da, which has no sequence homology with any other anticoagulant reported so far.
Sun
J
J
Department of Medical Zoology, Mie University School of Medicine, Tsu, Japan.
Yamaguchi
M
M
Yuda
M
M
Miura
K
K
Takeya
H
H
Hirai
M
M
Matsuoka
H
H
Ando
K
K
Watanabe
T
T
Suzuki
K
K
Chinzei
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D000925:Anticoagulants / Q000302:isolation & purification*; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification*; D006420:Hemeproteins / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D012239:Rhodnius / Q000737:chemistry*; D012469:Salivary Glands / Q000737:chemistry*; D012471:Salivary Proteins and Peptides / Q000235:genetics / Q000302:isolation & purification*; D017386:Sequence Homology, Amino Acid
D000925:Anticoagulants; D018076:DNA, Complementary; D006420:Hemeproteins; D012471:Salivary Proteins and Peptides; C090560:nitrophorin