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d -Ribulose 1,5-Diphosphate Carboxylase from Rhodospirillum rubrum

Journal of Biological Chemistry 1974年第11期249卷 3453-3458页

作者： F. Robert Tabita Bruce A. McFadden From the Graduate Program in Biochemistry and Department of Chemistry Washington State University Pullman Washington 99163

Ribulose 1,5-diphosphate carboxylase has been purified 15-fold from derepressed butyrate-grown Rhodospirillum rubrum to a specific activity of 1.76 µmoles of ribulose 1,5-diphosphate carboxylated per min per mg of protein at pH 8.0 and 30°. The enzyme, which is recovered in 30 to 35% yield, is homogeneous by the criterion of electrophoresis in gels polymerized from several concentrations of acrylamide. Immunochemical procedures were also used to establish homogeneity. The carboxylase is strongly activated by EDTA and dithiothreitol after storage if Mg 2+ is present. There is an absolute requirement of Mg 2+ for catalysis as only 0.05% activity is detected in its absence. Other divalent cations cannot be substituted for magnesium. Preincubation with Mg 2+ plus bicarbonate and initiation of catalysis with ribulose 1,5-diphosphate is required to achieve maximal rates. Homogeneous preparations could be stored without activity loss in the frozen state or at 2° in either phosphate or Tris buffers.

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Evidence for myosin-like intermediates in the hydrolysis of adenosine triphosphate by sperm-tail flagella.

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Journal of Biological Chemistry 1972年第12期247卷 4098-4100页

作者： Barclay, R. Yount, R.G. From the Graduate Program in Biochemistry Department of Chemistry Washington State University Pullman Washington 99163

The ATPase of glycerinated sperm flagella from the sea urchin ( Strongylocentrotus franciscanus ) catalyzes the preferential exchange of water oxygens into the phosphate originating from ATP. This type of exchange, so far known only for myosin-related ATPases, occurred under conditions (60 m m Mg 2+ , 1 to 10 m m Ca 2+ ) in which no inorganic phosphate-water exchange or exchange into unhydrolyzed ATP was observed. These results suggest that similar intermediates are formed by both skeletal myosin and flagellar ATPase during the cleavage of ATP, in spite of the known large differences in physical and enzymatic properties of the two contractile systems.

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The Kinetics and Mechanism of Fe(III) Exchange between Chelates and Transferrin

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Journal of Biological Chemistry 1967年第19期242卷 4284-4286页

作者： Carolyn Billups Leon Pape Paul Saltman From the Graduate Program of Biochemistry University of Southern California Los Angeles California 90007

With the use of Sephadex gel and collodion membrane filtrations it has been possible to determine the fraction of iron-ethylenediaminetetraacetate which reacts rapidly with apotransferrin. The values obtained for ratios of iron-EDTA to protein binding site of 1:1 (4 to 7% of binding sites occupied) and 5:1 (37% of binding sites occupied) correspond with similar values obtained by spectrophotometry. The suggested participation of a chemically active iron-EDTA species in the reaction is further strengthened.

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The Kinetics and Mechanism of Iron(III) Exchange between Chelates and Transferrin

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Journal of Biological Chemistry 1967年第12期242卷 2816-2821页

作者： George W. Bates Carolyn Billups Paul Saltman From the Graduate Program of Biochemistry University of Southern California Los Angeles California 90007

The rate of exchange of iron from ethylenediaminetetraacetate has been studied. The substance initially formed has a spectral absorption maximum at 520 mµ. In time, this peak shifts to 470 mµ, characteristic of the transferrin-iron complex. These results suggest the initial formation of an EDTA-iron-transferrin complex. Kinetic studies as a function of ferric EDTA and of apotransferrin concentrations are consistent with such a mechanism. The rate of removal of iron from ferric transferrin was in the order EDTA > citrate > nitrilotriacetic acid. The latter two chelates, however, reached equilibrium more rapidly. The importance of steric effects, as well as stability constants, in the biological utilization of trace metals is emphasized.

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EPITHELIAL-MESENCHYMAL INTERACTIONS RELATED TO PERIODONTAL DISEASE

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JOURNAL OF PERIODONTOLOGY 1970年第7期41卷 373-+页

作者： SLAVKIN, HC Department of Biochemistry School of Dentistry and Graduate Program of Cellular and Molecular Biology University of Southern California Los Angeles California 90007. The author is recipient of a Research Career Development Award from the National Institutes of Health.

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