Isolation from opossum serum of a metalloproteinase inhibitor homologous to human alpha 1B-glycoprotein.
Fractionation of opossum (Didelphis virginiana) serum with (NH4)2SO4, followed by chromatography on DEAE-Sepharose, phenyl-Sepharose, and Mono Q HR 5/5, has resulted in the isolation in homogeneous condition of a metalloproteinase inhibitor designated oprin (opossum proteinase inhibitor). Oprin is a single-chain glycoprotein (26% carbohydrate) with an estimated Mr = 52,000, pI = 3.5, and E(1%/1 cm) = 11. Oprin inhibited snake venom metalloproteinases, but showed no activity on venom serine proteinases or on bacterial metalloproteinases. Incubation of Crotalus atrox alpha-proteinase (EC 3.4.24.1) with oprin, and analysis of the reaction products by chromatography on Mono Q HR 5/5 and by electrophoresis under nondenaturing conditions, indicated formation of an inactive enzyme/inhibitor complex. The complex dissociated during SDS/polyacrylamide gel electrophoresis. An opossum liver cDNA library was immunoscreened, and clones containing cDNA encoding for part of the open reading frame for oprin were isolated. The cDNA inserts contained nucleotide sequences corresponding to two internal amino acid sequences of oprin which had been separately determined by protein sequence analysis. Protein database screening using a 211 amino acid sequence deduced from one of the cDNA inserts showed no significant homology to known proteinase inhibitors. There was, however, a 36% identity with human alpha 1B-glycoprotein, a plasma protein of unknown function related to the immunoglobulin supergene family. In addition, the amino-terminal sequence of oprin showed 46% identity with human alpha 1B-glycoprotein in a 26 amino acid residue overlap.(ABSTRACT TRUNCATED AT 250 WORDS)
Catanese
J J
JJ
Molecular and Cellular Biology Department, Roswell Park Cancer Institute, Buffalo, New York 14263.
Kress
L F
LF
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001798:Blood Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D003435:Crotalid Venoms / Q000037:antagonists & inhibitors; D004247:DNA / Q000302:isolation & purification; D006023:Glycoproteins; D006736:Horses; D006801:Humans; D007136:Immunoglobulins; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors*; D008969:Molecular Sequence Data; D009893:Opossums; D012689:Sequence Homology, Nucleic Acid; D013552:Swine
C048566:A1BG protein, human; D001798:Blood Proteins; D003435:Crotalid Venoms; D006023:Glycoproteins; D007136:Immunoglobulins; C032814:hemorrhagin components (Crotalid); C072438:oprin; D004247:DNA; D008666:Metalloendopeptidases