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Full-Text Articles in Biochemistry, Biophysics, and Structural Biology
Rhodobacter Capsulatus Porphobilinogen Synthase, A High Activity Metal Ion Independent Hexamer, David W. Bollivar, Cheryl Clauson, Rachel Lighthall, Siiri Forbes, Bashkim Kokona, Robert Fairman, Lenka Kundrat, Eileen K. Jaffe
Rhodobacter Capsulatus Porphobilinogen Synthase, A High Activity Metal Ion Independent Hexamer, David W. Bollivar, Cheryl Clauson, Rachel Lighthall, Siiri Forbes, Bashkim Kokona, Robert Fairman, Lenka Kundrat, Eileen K. Jaffe
David Bollivar
Background: The enzyme porphobilinogen synthase (PBGS), which is central to the biosynthesis of heme, chlorophyll and cobalamins, has long been known to use a variety of metal ions and has recently been shown able to exist in two very different quaternary forms that are related to metal ion usage. This paper reports new information on the metal ion independence and quaternary structure of PBGS from the photosynthetic bacterium Rhodobacter capsulatus. Results: The gene for R. capsulatus PBGS was amplified from genomic DNA and sequencing revealed errors in the sequence database. R. capsulatus PBGS was heterologously expressed in E. coli and …
Embryonic Expression Of Pre-Initiation Dna Replication Factors In Xenopus Laevis, Brian Walter, Jonathan Henry
Embryonic Expression Of Pre-Initiation Dna Replication Factors In Xenopus Laevis, Brian Walter, Jonathan Henry
Brian Walter
We examined the expression of various DNA replication factors, including: cdc45, the factors of the GINS heterotetramer (Sld5, Psf1, Psf2, Psf3), and PCNA, in Xenopus laevis during embryonic development via whole mount in situ hybridization. For the most part, these factors were expressed in similar patterns, with some subtle variations, throughout development within the anterior CNS, pharyngeal arches, and various placodes. More significant variations were also observed, including expression of only Psf1 and Psf2 in the pronephros and unique Psf2 expression in the somitic mesoderm. Overall, these results suggest that common regulatory mechanisms are involved in the transcriptional deployment of …
Thermal And Sodium Dodecylsulfate Induced Transitions Of Streptavidin, Mark Waner, Irina Navrotskaya, Amanda Bain, Edward Oldham, David Mascotti
Thermal And Sodium Dodecylsulfate Induced Transitions Of Streptavidin, Mark Waner, Irina Navrotskaya, Amanda Bain, Edward Oldham, David Mascotti
David P. Mascotti
The strong specific binding of streptavidin (SA) to biotin is utilized in numerous biotechnological applications. The SA tetramer is also known to exhibit significant stability, even in the presence of sodium dodecylsulfate (SDS). Despite its importance, relatively little is known about the nature of the thermal denaturation pathway for SA. This work uses a homogeneous SA preparation to expand on the data of previous literature reports, leading to the proposal of a model for temperature induced structural changes in SA. Temperature dependent data were obtained by SDS and native polyacrylamide gel electrophoresis (PAGE), differential scanning calorimetry (DSC), and fluorescence and …
Protein S-Glutathionylation In Retinal Pigment Epithelium Converts Heat Shock Protein 70 To An Active Chaperone., George Hoppe, Yuh-Cherng Chai, J. Crabb, Jonathan Sears
Protein S-Glutathionylation In Retinal Pigment Epithelium Converts Heat Shock Protein 70 To An Active Chaperone., George Hoppe, Yuh-Cherng Chai, J. Crabb, Jonathan Sears
Yuh-Cherng Chai
A disulfide bond between key redox-sensitive cysteine residues and glutathione is one mechanism by which redox related allosteric effectors can regulate protein structure and function. Here we test the hypothesis that glutaredoxin-1 (Grx-1), a member of the oxidoreductase family of enzymes, may be a critical component of redox-sensitive molecular switches by mediating reversible protein S-glutathionylation and enzymatic catalysis of thiol/disulfide exchange. Deglutathionylation of a 70 kDa protein by Grx-1 was detected using a monoclonal antibody specific to protein S-glutathionylation. Heat shock cognate protein 70 (Hsc70) was identified as a substrate of Grx-1 through mass spectrometry. Recombinant Hsc70 was glutathionylated in …
Molecular Profiling: Gene Expression Reveals Discrete Phases Of Lens Induction And Development In Xenopus Laevis, Brian Walter, Yimin Tian, Amy Garlisch, Maria Carinato, Matthew Elkins, Adam Wolfe, Jonathan Schaefer, Kimberly Perry, Jonathan Henry
Molecular Profiling: Gene Expression Reveals Discrete Phases Of Lens Induction And Development In Xenopus Laevis, Brian Walter, Yimin Tian, Amy Garlisch, Maria Carinato, Matthew Elkins, Adam Wolfe, Jonathan Schaefer, Kimberly Perry, Jonathan Henry
Brian Walter
No abstract provided.
Quantitative Nonisotopic Nitrocellulose Filter Binding Assays: Bacterial Manganese Superoxide Dismutase–Dna Interactions., Joshua Czerwinski, Stephanie Hovan, David Mascotti
Quantitative Nonisotopic Nitrocellulose Filter Binding Assays: Bacterial Manganese Superoxide Dismutase–Dna Interactions., Joshua Czerwinski, Stephanie Hovan, David Mascotti
David P. Mascotti
Nitrocellulose filter binding assays (NCFBAs) have been used for many years to qualitatively and quantitatively determine protein–nucleic acid affinities. While this technique can be robust thermodynamically and fairly simple to perform, the requirement of radiolabeled nucleic acids (typically 32P) has several major drawbacks. Some disadvantages are the short half-life of 32P, the inherent safety concerns, and the cost of working with radioisotopes. Another drawback is that over time the beta emissions cause fragmentation of the nucleic acids. We have modified standard NCFBAs by developing a quantitative nonisotopic chemiluminescent method using biotin-labeled DNA and a dual-filter format. The biotin tag is …