title: Molecules at the external nuclear surface. Sialic acid of nuclear membranes and electrophoretic mobility of isolated nuclei and nucleoli.
abstract: The molecules occurring as terminal residues on the external surfaces of nuclei prepared from rat liver by either sucrose-CaCl(2) or citric acid methods and nucleoli derived from the sucrose-CaCl(2) nuclei were studied chemically and electrokinetically. In 0.0145 M NaCl, 4.5% sorbitol, and 0.6 mM NaHCO(3) with pH 7.2 +/- 0.1 at 25 degrees C, the sucrose-CaCl(2) nuclei had an electrophoretic mobility of -1.92 microm/s/V/cm, the citric acid nuclei, -1.63 microm/s/V/cm, and the nucleoli, -2.53 microm/s/V/cm. The citric acid nuclei and the nucleoli contained no measurable sialic acid. The sucrose-CaCl(2) nuclei contained 0.7 nmol of sialic acid/mg nuclear protein; this was essentially located in the nuclear envelope. Treatment of these nuclei with 50 microg neuraminidase/mg protein resulted in release of 0.63 nmol of sialic acid/mg nuclear protein; treatment with 1 % trypsin caused release of 0.39 nmol of the sialic acid/mg nuclear protein. The pH-mobility curves for the particles indicated the sucrose-CaCl(2) nuclei surface had an acid-dissociable group of pK. approximately 2.7 while the pK for the nucleoli was considerably lower. Nucleoli treated with 50 microg neuraminidase/mg particle protein had a mobility of -2.53 microm/s/V/cm while sucrose-CaCl(2) nuclei similarly treated had a mobility of -1.41 microm/s/V/cm. Hyaluronidase at 50 microg/mg protein had no effect on nucleoli mobility but decreased the sucrose-CaCl(2) nuclei mobility to -1.79 microm/s/V/cm. Trypsin at 1 % elevated the electrophoretic mobility of the sucrose-CaCl(2) nuclei slightly but decreased the mobility of the nucleoli to -2.09 microm/s/V/cm. DNase at 50 microg/mg protein had no effect on the mobility of the isolated sucrose-CaCl(2) nuclei but decreased the electrophoretic mobility of the nucleoli to -1.21 microm/s/V/cm. RNase at 50 microg/mg protein also had no effect on the electrophoretic mobility of the sucrose-CaCl(2) nuclei but decreased the nucleoli mobility to -2.10 microm/s/V/cm. Concanavalin A at 50 microg/mg protein did not alter the nucleoli electrophoretic mobility but decreased the sucrose-CaCl(2) nuclei electrophoretic mobility to -1.64 microm/s/V/cm. The results are interpreted to mean that the sucrose-CaCl(2) nuclear external surface contains terminal sialic acid residues in trypsin-sensitive glycoproteins, contains small amounts of hyaluronic acid, is completely devoid of nucleic acids, and binds concanavalin A. The nucleolus surface is interpreted to contain a complex made up of protein, RNA, and primarily DNA, to be devoid of sialic acid and hyaluronic acid, and not to bind concanavalin A.
mesh_terms: D000818:Animals; D002122:Calcium Chloride / Q000494:pharmacology; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D002462:Cell Membrane / Q000032:analysis*; D002466:Cell Nucleolus / Q000032:analysis*; D002467:Cell Nucleus / Q000032:analysis*; D002951:Citrates / Q000494:pharmacology; D003208:Concanavalin A / Q000494:pharmacology; D004247:DNA / Q000032:analysis; D003851:Deoxyribonucleases / Q000494:pharmacology; D004586:Electrophoresis; D004912:Erythrocytes / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis; D006821:Hyaluronoglucosaminidase / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000166:cytology; D008853:Microscopy; D009439:Neuraminidase / Q000494:pharmacology; D011506:Proteins / Q000032:analysis; D012313:RNA / Q000032:analysis; D051381:Rats; D012260:Ribonucleases / Q000494:pharmacology; D013395:Sucrose / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology; D014361:Trypsin Inhibitors / Q000494:pharmacology; D014465:Ultrasonics
chemical_list: D002241:Carbohydrates; D002951:Citrates; D006023:Glycoproteins; D011506:Proteins; D014361:Trypsin Inhibitors; D003208:Concanavalin A; D013395:Sucrose; D012313:RNA; D006820:Hyaluronic Acid; D004247:DNA; D003851:Deoxyribonucleases; D012260:Ribonucleases; D009439:Neuraminidase; D006821:Hyaluronoglucosaminidase; D014357:Trypsin; D002122:Calcium Chloride