Characterization of the antihemorrhagic factors of mongoose (Herpestes edwardsii).
Three antihemorrhagic factors (AHF1, AHF2 and AHF3) isolated from the serum of mongoose (Herpestes edwardsii) are glycoproteins of monomer structure with the same mol. wt (about 65,000), which contain 4.2%, 13.6% and 6.0% carbohydrates as glucose, respectively. All are composed of about 600 amino acids of similar composition. The 32 amino terminal amino acid sequences of three antihemorrhagic factors were determined, and sequence homologies were examined. AHF1 and AHF2 were of the same amino acid sequence, and showed high homologies to AHF3, oprin (opossum proteinase inhibitor) and human alpha 1B-glycoprotein; 68.7%, 42.3% and 50.0% identity, respectively. AHF1 completely inhibited the hemorrhagic activity of HR2b, the hemorrhagic factor of habu snake, at the concentration of five-fold molar excess, although incomplete inhibition (50%) of proteinase activity of the hemorrhagic factor was observed even at the concentration of 20-fold molar excess of antihemorrhagic factor. Incubation of HR2b with AHF1, and analysis of the reaction products by chromatography on TSK gel G-3000SW and on the ultracentrifuge did not show formation of an inactive enzyme inhibitor complex. However, the complex formation between AHF1 and HR2b was observed by a BIAcore analysis and TSK gel SP-5PW column chromatography. No alteration in the primary or the secondary structure of both factors was demonstrated by SDS-PAGE and circular dichroism spectrum at the far-UV wavelength before and after incubation of both factors, respectively.
Qi
Z Q
ZQ
Department of Bioscience and Biotechnology, University of The Ryukyus, Okinawa, Japan.
Yonaha
K
K
Tomihara
Y
Y
Toyama
S
S
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis / Q000378:metabolism; D000818:Animals; D000997:Antivenins / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D001798:Blood Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000032:analysis; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003435:Crotalid Venoms / Q000037:antagonists & inhibitors / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006470:Hemorrhage / Q000188:drug therapy; D008988:Herpestidae; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011817:Rabbits; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012910:Snake Venoms / Q000037:antagonists & inhibitors; D014461:Ultracentrifugation
D000596:Amino Acids; D000997:Antivenins; D001798:Blood Proteins; D002241:Carbohydrates; D003435:Crotalid Venoms; D012910:Snake Venoms; C003941:antihemorrhagic factor; C052578:hemorrhagic toxin HR-2b