Characterization and sequencing of rabbit, pig and mouse angiogenins: discernment of functionally important residues and regions.
Rabbit, pig and mouse angiogenins have been purified from blood serum and characterized, and the rabbit and pig proteins have been sequenced fully. A partial sequence of the mouse protein is consistent with the sequence deduced from the genomic DNA (Bond, M.D. and Vallee, B.L. (1990) Biochem. Biophys. Res. Commun. 171, 988-995). All three angiogenins are homologous to the pancreatic RNases and contain the essential catalytic residues His-13, Lys-40 and His-114, and the 6 half-cystines of the human protein. Like human angiogenin they display extremely low ribonucleolytic activities toward wheat-germ RNA, yeast RNA, poly(C) and poly(U). The rabbit and pig proteins induce neovascularization in vivo and also inhibit protein synthesis in vitro. The interaction of rabbit, pig and bovine angiogenins with placental ribonuclease inhibitor, a potent inhibitor of angiogenin, was examined by fluorescence spectroscopy. Rate and equilibrium binding constants indicate that rabbit angiogenin binds to the inhibitor much like human angiogenin, whereas the pig and bovine proteins show significant differences. A comparison of the five angiogenin sequences now available points to specific residues that are highly conserved among them but differ from the corresponding residues in the RNases. These residues are clustered in particular regions of the three-dimensional structure, two of which contribute to the angiogenic, second-messenger and/or protein synthesis inhibition activities of human angiogenin.
Bond
M D
MD
Center for Biochemical and Biophysical Sciences and Medicine, Harvard Medical School, Boston, MA 02115.
Strydom
D J
DJ
Vallee
B L
BL
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010926:Placental Hormones / Q000737:chemistry*; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D011817:Rabbits; D012259:Ribonuclease, Pancreatic; D016415:Sequence Alignment; D013552:Swine; D014364:Tryptophan / Q000032:analysis*
D000596:Amino Acids; D010926:Placental Hormones; D011506:Proteins; C022052:placental ribonuclease inhibitor; D014364:Tryptophan; C046366:angiogenin; D012259:Ribonuclease, Pancreatic