title: Primary structure of hemorrhagic protein, HR2a, isolated from the venom of Trimeresurus flavoviridis.
abstract: The complete amino acid sequence and disulfide bridge location of HR2a, one of the hemorrhagic proteins isolated from the snake venom of Trimeresurus flavoviridis, have been determined by analysis of peptides derived from digests with cyanogen bromide, lysyl endopeptidase, trypsin, and Staphylococcus aureus V8 protease. Peptides were purified by gel filtration followed by reversed-phase HPLC. HR2a has the amino-terminal sequence of less than Glu-Gln-Arg- and consists of a total of 202 residues with a calculated molecular weight of 23,015. Sequence analysis indicates the presence of another isoform which lacks the amino-terminal residue, making 201 amino acid residues with a molecular weight of 22,887. Three disulfide bridges of HR2a link Cys-118 to Cys-197, Cys-159 to Cys-181, and Cys-161 to Cys-164. HR2a contains a segment which is similar to the zinc-chelating sequences found in thermolysin and several mammalian metalloproteinases, suggesting that HR2a is a metalloproteinase with limited substrate specificity. However, there is no other significant sequence homology with thermolysin except for the zinc-ligand region.
authors.affiliation: Department of Biology, Faculty of Science, Kyushu University, Fukuoka.
authors.forename: T
authors.initials: T
authors.lastname: Miyata
authors.forename: H
authors.initials: H
authors.lastname: Takeya
authors.forename: Y
authors.initials: Y
authors.lastname: Ozeki
authors.forename: M
authors.initials: M
authors.lastname: Arakawa
authors.forename: F
authors.initials: F
authors.lastname: Tokunaga
authors.forename: S
authors.initials: S
authors.lastname: Iwanaga
authors.forename: T
authors.initials: T
authors.lastname: Omori-Satoh
mesh_terms: D000595:Amino Acid Sequence; D003435:Crotalid Venoms / Q000032:analysis*; D004220:Disulfides / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis / Q000302:isolation & purification; D011487:Protein Conformation; D012697:Serine Endopeptidases; D013438:Sulfhydryl Compounds / Q000032:analysis; D014357:Trypsin
chemical_list: D003435:Crotalid Venoms; D004220:Disulfides; D010455:Peptides; D013438:Sulfhydryl Compounds; C052577:hemorrhagic toxin HR-2a; D012697:Serine Endopeptidases; D014357:Trypsin; C036813:lysyl endopeptidase