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16 results on '"Speight, Lee"'

1. Sustainable sustainability claims

Author

Speight, Lee Colyer, Lanci, Christopher J., and Mazeffa, Douglas

Subjects
Sustainable development, Raw materials, Green technology, Business, Chemicals, plastics and rubber industries
Abstract

Greenwashing, or the practice of misrepresenting or over representing the environmental benefits of a product or practice, is common in consumer marketing. The term was coined in 1986 by Jay [...]

Published
2015

2. Everything old is new again

Author

Speight, Lee Colyer

Subjects
Raw materials, Adhesives and sealants industry, Petroleum chemicals, Business, Chemicals, plastics and rubber industries
Abstract

This is a call for innovation and collaboration. The ascent--or, more appropriately--re-ascent of renewable chemistry provides great promise for the adhesives and sealants industry. However, the full promise of these [...]

Published
2015

5. Improving target amino acid selectivity in a permissive aminoacyl tRNA synthetase through counter-selection

Author

Sungwienwong, Itthipol, primary, Hostetler, Zachary M., additional, Blizzard, Robert J., additional, Porter, Joseph J., additional, Driggers, Camden M., additional, Mbengi, Lea Z., additional, Villegas, José A., additional, Speight, Lee C., additional, Saven, Jeffery G., additional, Perona, John J., additional, Kohli, Rahul M., additional, Mehl, Ryan A., additional, and Petersson, E. James, additional

Published
2017
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6. Efficient Synthesis and In Vivo Incorporation of Acridon-2-ylalanine, a Fluorescent Amino Acid for Lifetime and FRET/LRET Studies

Author

Speight, Lee C., Muthusamy, Anand K., Goldberg, Jacob M., Warner, John B., Wissner, Rebecca F., Willi, Taylor S., Woodman, Bradley F., Mehl, Ryan A., and Petersson, E. James

Subjects
Models, Molecular, Alanine, Luminescence, Fluorescence Resonance Energy Transfer, Acridines, Article
Abstract

The amino acid acridon-2-ylalanine (Acd) can be a valuable probe of protein conformational change because it is a long lifetime, visible wavelength fluorophore that is small enough to be incorporated during ribosomal biosynthesis. Incorporation of Acd into proteins expressed in Escherichia coli requires efficient chemical synthesis to produce large quantities of the amino acid and the generation of a mutant aminoacyl tRNA synthetase that can selectively charge the amino acid onto a tRNA. Here, we report the synthesis of Acd in 87% yield over five steps from Tyr and the identification of an Acd synthetase by screening candidate enzymes previously evolved from Methanococcus janaschii Tyr synthetase for unnatural amino acid incorporation. Furthermore, we characterize the photophysical properties of Acd, including quenching interactions with select natural amino acids and Förster resonance energy transfer (FRET) interactions with common fluorophores such as methoxycoumarin (Mcm). Finally, we demonstrate the value of incorporation of Acd into proteins, using changes in Acd fluorescence lifetimes, Mcm/Acd FRET, or energy transfer to Eu(3+) to monitor protein folding and binding interactions.

Published
2013

8. Minimalistic probes for protein folding and structure studies

Author

Speight, Lee Colyer and Speight, Lee Colyer

Abstract

Site-specific incorporation of fluorescent probes into proteins enhances the ability of researchers to study protein folding and structural dynamics. Currently, the most common fluorescent probe for proteins is green fluorescent protein (GFP). However, the use of GFP, a 28 kDa protein, requires its covalent attachment to a protein of interest. Additionally, this attachment is restricted to either the N- or C-terminus of the protein, drastically limiting the scope of experiments possible. Alternative technologies for the addition of novel fluorescence properties to proteins exist, however they are also limited by the probe size or its method of attachment. Since many proteins of interest are less than or equal to the size of these fluorescent auxiliaries, it is likely that the covalent fusion of the fluorescent apparatus will significantly perturb the native fold or function of the molecule of interest. As such, the development of minimalistic probes for protein functional studies is of great interest to the biochemical community. Acridon-2-ylalanine (Acd) is a minimalistic fluorescent unnatural amino acid (flUAA) that possess a long lifetime, resistance to photobleaching, a near unity quantum yield, and visible wavelength emissions. While there existed literature precedent for the synthesis of Acd, these methods required harsh conditions and reagents that precluded a high-yielding, scalable synthesis of the flUAA. To this end, we have devised an efficient, scalable 5-step synthesis utilizing Tyr that yields Acd in 87 % overall yield. Recently, we have shown that Acd can be incorporated into proteins in vivo and can be a valuable probe of protein-peptide interactions and protein conformational change due to its blue-wavelength excitation, unique solvatochromic properties, and ability to participate in energy transfer with endogenous amino acids (Trp and Tyr), exogenous fluorophores (methoxycoumarin), and the lanthanide ion Eu 3+. Furthermore, as the need for minimali

Published
2014

11. Efficient Synthesis and In Vivo Incorporation of Acridon-2-ylalanine, a Fluorescent Amino Acid for Lifetime and Förster Resonance Energy Transfer/Luminescence Resonance Energy Transfer Studies

Author

Speight, Lee C., primary, Muthusamy, Anand K., additional, Goldberg, Jacob M., additional, Warner, John B., additional, Wissner, Rebecca F., additional, Willi, Taylor S., additional, Woodman, Bradley F., additional, Mehl, Ryan A., additional, and Petersson, E. James, additional

Published
2013
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15. Sustainable Sustainability Claims.

Author

Colyer Speight, Lee, Lanci, Christopher J., and Mazeffa, Douglas

Subjects
GREENWASHING (Marketing), COATINGS industry, SURFACE coatings, RAW materials, MATERIALS
Abstract

The article focuses on the controversy over greenwashing of coatings and its impact on the industry. Topics discussed include greenwashing refers to misrepresenting the environmental advantages of a product or habit, a standard rule for assessing environmental effects of coatings applies on 10 percent for formulation, 40 percent for downstream usage, and 50 percent for raw material production, and sustainable raw material sourcing methods like book and claim, and mass balance.

Published
2015

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