Two distinct secretory ribonucleases from human cerebrum: purification, characterization and relationships to other ribonucleases.
Two RNAases from human cerebrum were purified to an electrophoretically homogeneous state and their molecular masses were 22.0 kDa (tentatively called RNAase HB-1) and 19.0 kDa (RNAase HB-2). Analyses of the amino acid compositions, N-terminal amino acid sequences and catalytic properties of these enzymes provided strong evidence that they were strictly related to the secretory (sec) RNAases, such as the pancreatic enzyme, very similar immunologically to urinary sec RNAase, but clearly distinguishable from urinary non-secretory (nonsec) RNAase. There were several differences between HB-1 and HB-2, namely their immunological reactivities with specific antibodies, heat-stabilities, attached carbohydrate moieties and molecular masses. In particular, HB-2 appeared to be nonglycosylated, in view of its lack of affinity for several conjugated lectins, the absence of hexosamine and no change in electrophoretic mobility before and after peptide:N-glycosidase F digestion, whereas HB-1 and human sec RNAases purified from kidney, pancreas and urine all appeared to be glycosylated, as they moved to the same position as HB-2 when electrophoresed after glycosidase digestion. An antibody against urinary sec RNAase inhibited 75% and 20% of the total activity of the crude cerebral extract against RNA at pH 8.0 and 6.0 respectively, whereas an antibody against urinary nonsec RNAase had no such inhibitory effect. These findings suggest that yet another type(s) of cerebral RNAase, which is unable to cross-react immunologically with sec and nonsec RNAases, may exist. Two RNAases corresponding to HB-1 and HB-2 were identified in fresh cerebrospinal fluid.
Yasuda
T
T
Department of Legal Medicine, Fukui Medical School, Japan.
Nadano
D
D
Takeshita
H
H
Kishi
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology*; D050260:Carbohydrate Metabolism; D002384:Catalysis; D002417:Cattle; D002555:Cerebrospinal Fluid / Q000201:enzymology; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007668:Kidney / Q000201:enzymology; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology; D010926:Placental Hormones / Q000494:pharmacology; D012260:Ribonucleases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013154:Spleen / Q000201:enzymology
D010926:Placental Hormones; C022052:placental ribonuclease inhibitor; D012260:Ribonucleases