A5012029286- Bacterial Genetics and Biotechnology
- DNA and Nucleic Acid Chemistry
- RNA and protein synthesis mechanisms
- Microfluidic and Capillary Electrophoresis Applications
- Enzyme Structure and Function
- Alkaline Phosphatase Research Studies
- Biochemical and Molecular Research
- Hemoglobin structure and function
- Growth Hormone and Insulin-like Growth Factors
- Microbial Metabolic Engineering and Bioproduction
- Mass Spectrometry Techniques and Applications
- Protein Structure and Dynamics
Beloit College
2012-2017
University of Wisconsin–Madison
2008-2012
Though opening of the start site (+1) region promoter DNA is required for transcription by RNA polymerase (RNAP), surprisingly little known about how and when this occurs in mechanism. Early events at lambdaP(R) load duplex into active cleft Escherichia coli RNAP, forming closed, permanganate-unreactive intermediate I(1). Conversion to subsequent I(2) overcomes a large enthalpic barrier. Is open? Here we create burst rapidly destabilizing open complexes (RP(o)) with 1.1 M NaCl. Fast...
Transcription by all RNA polymerases (RNAPs) requires a series of large-scale conformational changes to form the transcriptionally competent open complex RP(o). At lambdaP(R) promoter, Escherichia coli sigma(70) RNAP first forms wrapped, closed 100 bp I(1). The subsequent step opens entire DNA bubble, creating relatively unstable (open) I(2). Additional convert I(2) stable Here we probe these events dissecting effects Na(+) salts Glu(-), F(-), and Cl(-) on each in this critical process....
Differences in kinetics of transcription initiation by RNA polymerase (RNAP) at different promoters tailor the pattern gene expression to cellular needs. After initial binding, large conformational changes occur promoter DNA and RNAP form initiation-capable complexes. To understand mechanism regulation initiation, nature sequence these must be determined. Escherichia coli uses binding free energy unwind separate 13 base pairs λPR unstable open intermediate I2, which rapidly converts much...
The three-dimensional nature of interactions between enzymes and their substrates often leads to exacting spatial binding orientations stereoselectivity in chemical catalysis. Dehydrogenases that use NAD+ as a redox cofactor tend show stereospecificity transferring hydride the C4 nicotinamide moiety via either re or si face. this transfer, which results prochiral reduced NADH, may be determined using deuterated 1H NMR spectroscopy. A biochemistry lab activity combines analysis intermolecular...
The interaction between oxygen and hemoglobin is a classic example of cooperative ligand-binding process. This gaseous-ligand binding process may be studied in the undergraduate laboratory with variable-pressure cuvette apparatus. An updated method described to assemble apparatus composed inexpensive supplies equipment commonly found chemistry laboratory.
Alkaline phosphatase catalyzes the hydrolysis of phosphate ester bonds, which results in removal from many biologically relevant molecules (i.e. DNA, proteins, carbohydrates). This enzyme functions a wide variety organisms ranging bacteria to mammals and has vitro applications for genetic engineering. Numerous kinetic crystal structure studies over past fifty years have revealed details alkaline catalytic mechanism. However, few investigated solution‐phase conformational changes that...