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Articles 1 - 9 of 9

Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Purification, Surfactant Stabilization, Molecular Weight Studies, And Divalent Metal Ion Kinetics Of Dextransucrase From Leuconostoc Mesenteroides Nrrl B-512f , Arthur West Miller Jan 1985

Purification, Surfactant Stabilization, Molecular Weight Studies, And Divalent Metal Ion Kinetics Of Dextransucrase From Leuconostoc Mesenteroides Nrrl B-512f , Arthur West Miller

Retrospective Theses and Dissertations

Dextransucrase from Leuconostoc mesenteroides NRRL B-512F, an enzyme that synthesizes an (alpha)-(1(--->)6)-linked glucan with (alpha)-(1(--->)3)-linked branches from the glucosyl moiety of sucrose, was stabilized against thermal inactivation and against inactivation caused by dilution and manipulation by the addition of nonionic detergents, mild ionic detergents, poly(ethylene glycol), or high-molecular-weight dextran;Dextransucrase was purified by a sequence of dextranase treatment, DEAE-cellulose chromatography, affinity chromatography on Sephadex G-200, and DEAE-Trisacryl M chromatography. The purified enzyme had a specific activity of 170 U/mg protein and no detectable carbohydrate;The enzyme consisted of two forms of differing ...


Production And Use Of Monoclonal Antibodies For Molecular Characterization, Affinity Purification, And Serological Detection Of Economically Significant Plant Viruses , Robert Diaco Jan 1985

Production And Use Of Monoclonal Antibodies For Molecular Characterization, Affinity Purification, And Serological Detection Of Economically Significant Plant Viruses , Robert Diaco

Retrospective Theses and Dissertations

Hybridomas secreting monoclonal antibodies (M-Abs) against three isolates of barley yellow dwarf virus (BYDV) were established. Two M-Abs were generated against the MAV isolate, one against RPV, and six against P-PAV. None of the M-Abs reacted with healthy host components. Reactions of M-Abs, or unlabelled polyclonal antisera, in an indirect enzyme-linked immunosorbent assay (ELISA) indicate the presence of a common epitope on all three virus isolates. BYDV dissociated when incubated in carbonate-bicarbonate coating buffer at pH 9.6; stabilization was achieved by predialysis with either 2% formaldehyde or 2% glutaraldehyde. In indirect ELISA, unlabelled polyclonal antisera bound to both stabilized ...


Studies Of The Hela Mitochondrial Translation System , Thomas James Girard Jan 1985

Studies Of The Hela Mitochondrial Translation System , Thomas James Girard

Retrospective Theses and Dissertations

Mitochondrial (mt) ribosomes and ribosomal subunits were prepared from HeLa S(,3) (chloramphenicol (CAP) sensitive) and 296-1 (CAP resistant) cell lines and used in in vitro protein synthesis activity studies. Mt ribosomes isolated in low salt (100 mM KCl, 30 mM MgCl(,2)) were active in poly(U)-directed protein synthesis when supplemented with E. coli tRNA and E. coli supernatant fraction (S-100). Washing these ribosomes with high salt-containing buffers (500 mM KCl, 5 mM MgCl(,2)) results in the loss of protein synthesis activity. Activity by high salt washed mt ribosomes was restored by the addition of a mt ...


Synthesis, Analysis And Characterization Of Deuterated Analogues Of Vitamin A , Harold Robert Bergen Iii Jan 1985

Synthesis, Analysis And Characterization Of Deuterated Analogues Of Vitamin A , Harold Robert Bergen Iii

Retrospective Theses and Dissertations

Several deuterated analogues of vitamin A have been synthesized by two methods. The aldehyde, 20, 20, 20-('2)H(,3) retinaldehyde, was prepared by the approach of Tanis et al. (Tetrahedron Lett. 1978, 10, 869-872) based on a modification of Wittig's directed aldol condensation. Four more deuterated analogues of vitamin A were synthesized based on the use of a Wittig-Horner reaction and deuterium exchange for the incorporation of deuterium. These analogues included 20,20,20-('2)H(,3) retinal, 14,20,20,20-('2)H(,4) retinol, 12,14,20,20,20-('2)H(,5)-retinol and 10,19 ...


The Interaction Of Adenosine-5'-Monophosphate And Fructose 2,6-Bisphosphate With Bovine Liver Fructose 1,6-Bisphosphatase , Nancy Joan Ganson Jan 1985

The Interaction Of Adenosine-5'-Monophosphate And Fructose 2,6-Bisphosphate With Bovine Liver Fructose 1,6-Bisphosphatase , Nancy Joan Ganson

Retrospective Theses and Dissertations

The fructose 1,6-bisphosphatase (FBPase) reaction was investigated in the reverse direction by using fructose 2,6-bisphosphate (2,6FBP). The effector was found to be a potent inhibitor of the reverse reaction substrates fructose 6-phosphate (F6P) and inorganic phosphate (P(,i)). Inhibition of bovine liver FBPase by 2,6FBP was competitive with respect to both substrates, and slope replots were linear. In the context of other accumulated kinetic data, the results serve to support a Random Bi Uni mechanism as the most likely mechanism for the reverse reaction. In addition, two models consistent with the data are presented for the ...


6-Fluoropyridoxal Phosphate And Pyridoxal Sulfate In Aspartate Aminotransferase , Robert D. Scott Jan 1985

6-Fluoropyridoxal Phosphate And Pyridoxal Sulfate In Aspartate Aminotransferase , Robert D. Scott

Retrospective Theses and Dissertations

The chemical and spectroscopic properties of 6-fluoropyridoxal 5'-phosphate, of its Schiff base with valine and of 6-fluoropyridoxamine 5'-phosphate have been investigated. The modified coenzymes have also been combined with the apo form of cytosolic aspartate aminotransferase and the properties of the resulting enzymes and of their complexes with substrates and inhibitors have been recorded. Although the presence of the 6-fluoro substituent reduces the basicity of the ring nitrogen over 10,000-fold, the modified coenzymes bind predominately in their dipolar ionic forms as do the natural coenzymes. Enzyme containing the modified coenzymes binds substrates and dicarboxylate substrates normally, and ...


Studies On The Sucrose Binding Site Of The Glucosyltransferases Of Leuconostoc And Streptococcus Bacteria , Thomas Paul Binder Jan 1985

Studies On The Sucrose Binding Site Of The Glucosyltransferases Of Leuconostoc And Streptococcus Bacteria , Thomas Paul Binder

Retrospective Theses and Dissertations

para-Nitrophenyl-(alpha)-D-glucopyranoside was found to be a substrate for the glucosyltransferases produced by several species of Streptococcus and Leuconostoc bacteria. It was found to react to produce both high molecular weight dextran and acceptor products with the glucosyltransferase of Leuconostoc mesenteroide B512F;The glucosyltransferases were also found to react with alternate glucosyl donors such as dextran, maltotriose, panose, and isomaltodextrins containing three or more glucose units. These reactions seemed to be the reverse of acceptor reactions;In order to study the sucrose binding sites of these enzymes, techniques were developed for the synthesis of sucrose analogs. By these techniques ...


The Effect Of Diabetes And Insulin On The Turnover Of Hexokinase Ii In The Skeletal Muscle , Sandra Kay Frank Jan 1985

The Effect Of Diabetes And Insulin On The Turnover Of Hexokinase Ii In The Skeletal Muscle , Sandra Kay Frank

Retrospective Theses and Dissertations

Studies were undertaken to determine the nature of the cause(s) for the reported decrease in the measurable hexokinase II activity in the skeletal muscle of the streptozotocin-diabetic rat. An ELISA procedure was developed to determine the amount of hexokinase II protein in the skeletal muscle extracts, and immunoprecipitation was utilized to determine the hexokinase II activity. Both hexokinase II activity and the amount of hexokinase II protein decreased in the diabetic rat. Both increased toward normal treatment with insulin. The specific activity of the hexokinase II was found to be the same in the skeletal muscle extracts from normal ...


Subunit Structure Of The Protofilament Buidling Blocks Of Intermediate Filaments , Yuk-Ying Susana Pang Jan 1985

Subunit Structure Of The Protofilament Buidling Blocks Of Intermediate Filaments , Yuk-Ying Susana Pang

Retrospective Theses and Dissertations

The purposes of this study were to use chemical cross-linking reagents to elucidate the number of subunits in desmin and vimentin protofilaments, to elucidate conditions necessary for dissociation of the subunits of desmin protofilaments, and to understand the arrangement of the subunit chains of the protofilaments. Three chemical cross-linkers, namely ethylene glycolbis(succinimidyl succinate), bis 2-(succinimidooxy-carbonyloxy)ethyl sulfone, and glutaraldehyde, were used to examine the subunit composition of the approximately 2.5 nm diameter protofilaments that are present in pH 8.5, low ionic strength solutions of highly purified avian smooth muscle desmin or of mammalian smooth muscle vimentin ...