scholarly journals Purification and characterization of prolixin S (nitrophorin 2), the salivary anticoagulant of the blood-sucking bug Rhodnius prolixus

1995 ◽  
Vol 308 (1) ◽  
pp. 243-249 ◽  
Author(s):  
J M C Ribeiro ◽  
M Schneider ◽  
J A Guimarães
Keyword(s):  
Cation Exchange ◽  
Salivary Glands ◽  
Coagulation Factor ◽  
Rhodnius Prolixus ◽  
Exchange Chromatography ◽  
Factor X ◽  
Blood Sucking ◽  
Weak Cation Exchange ◽  
Haem Protein ◽  
Strong Cation Exchange Chromatography

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.

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

1996 ◽  
Vol 75 (04) ◽  
pp. 573-577 ◽  
Author(s):  
Jianxin Sun ◽  
Masahiro Yamaguchi ◽  
Masao Yuda ◽  
Ken Miura ◽  
Hiroyuki Takeya ◽  
...  
Keyword(s):  
Salivary Glands ◽  
Protein A ◽  
Gel Filtration ◽  
Specific Inhibitor ◽  
Rhodnius Prolixus ◽  
Amino Acid Sequences ◽  
Factor X ◽  
Blood Sucking ◽  
Red Color ◽  
Absorbance Peak

SummaryThe 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 Villa. 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.

Strong cation-exchange chromatography of proteins on a sulfoalkylated monolithic cryogel

2015 ◽  
Vol 1386 ◽  
pp. 13-21 ◽  
Author(s):  
Işık Perçin ◽  
Rushd Khalaf ◽  
Bastian Brand ◽  
Massimo Morbidelli ◽  
Orhan Gezici
Keyword(s):  
Cation Exchange ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Strong Cation Exchange ◽  
Strong Cation Exchange Chromatography

Characterization of the Acidic Species of a Monoclonal Antibody Using Weak Cation Exchange Chromatography and LC-MS

2015 ◽  
Vol 87 (17) ◽  
pp. 9084-9092 ◽  
Author(s):  
Gomathinayagam Ponniah ◽  
Adriana Kita ◽  
Christine Nowak ◽  
Alyssa Neill ◽  
Yekaterina Kori ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cation Exchange ◽  
Exchange Chromatography ◽  
Cation Exchange Chromatography ◽  
Weak Cation Exchange

scholarly journals Salivary apyrase of Rhodnius prolixus. Kinetics and purification

1986 ◽  
Vol 233 (3) ◽  
pp. 885-891 ◽  
Author(s):  
J J F Sarkis ◽  
J A Guimarães ◽  
J M C Ribeiro
Keyword(s):  
Atp Hydrolysis ◽  
Competitive Inhibition ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Rhodnius Prolixus ◽  
Exchange Chromatography ◽  
Phosphate Esters ◽  
Inorganic Pyrophosphatase ◽  
Blood Sucking ◽  
Specific Activities

The salivary apyrase activity of the blood-sucking bug Rhodnius prolixus was found to reside in a true apyrase (ATP diphosphohydrolase, EC 3.6.1.5) enzyme. The crude saliva was devoid of 5′-nucleotidase, inorganic pyrophosphatase, phosphatase and adenylate kinase activities. ATP hydrolysis proceeded directly to AMP and Pi without significant accumulation of ADP. Km values for ATP and ADP hydrolysis were 229 and 291 microM respectively. Ki values for ATP and ADP inhibition of ADP and ATP hydrolysis were not different from the Km values, and these experiments indicated competitive inhibition. Activities were purified 126-fold by combined gel filtration and ion-exchange chromatography procedures with a yield of 63%. The purified enzyme displayed specific activities of 580 and 335 mumol of Pi released/min per mg of protein for ATP and ADP hydrolysis respectively. The action of the purified enzyme on several phosphate esters indicates that Rhodnius apyrase is a non-specific nucleosidetriphosphate diphosphohydrolase.

scholarly journals Studies on Biological Active Compounds of the Salivary Glands in Blood Sucking Insects and Ticks : Anti-coagulants in Rhodnius prolixus

1996 ◽  
Vol 47 (2) ◽  
pp. 201
Author(s):  
Yasuo Chinzei ◽  
Jianxin Sun ◽  
Masao Yuda ◽  
Masahiro Yamaguchi ◽  
Ken Miura ◽  
...  
Keyword(s):  
Salivary Glands ◽  
Rhodnius Prolixus ◽  
Active Compounds ◽  
Blood Sucking
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