Abstract

The overall objective of this research is to gain greater insight into the structure, mechanism of action and metabolic control of acyl-CoA dehydrogenases. Inactive derivatives of the pig kidney acyl-CoA dehydrogenase with iodoacetic acid and several dione reagents are under investigation to identify catalytically essential amino acid residues. Two mechanism based inhibitors of the enzyme, methylenecylcopropylacetyl-CoA and 3,4-pentadienoyl-CoA, are under investigation and preliminary results are suggestive of a mechanism of substrate dehydrogenation involving carbanion formation. The mechanism of stabilization of the red, anionic, seiquinone by acyl-CoA derivatives is under investigation. The results should aid in the interpretation of the mechanism of action of these acyl-CoA dehydrogenases. We will extend our preliminary studies on the reconstitution of pig kidney general acyl-CoA dehydrogenase using ring modified flavin analogs. Efforts will continue to develop a satisfactory purification scheme for pig kidney electron transferring flavoprotein.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM026643-08
Application #
3274042
Study Section
Physical Biochemistry Study Section (PB)
Project Start
1979-07-01
Project End
1987-06-30
Budget Start
1986-07-01
Budget End
1987-06-30
Support Year
8
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Delaware
Department
Type
Schools of Arts and Sciences
DUNS #
059007500
City
Newark
State
DE
Country
United States
Zip Code
19716

  Publications

Hudson, Devin A; Caplan, Jeffrey L; Thorpe, Colin (2018) Designing Flavoprotein-GFP Fusion Probes for Analyte-Specific Ratiometric Fluorescence Imaging. Biochemistry 57:1178-1189
Yu, Tiantian; Laird, Joanna R; Prescher, Jennifer A et al. (2018) Gaussia princeps luciferase: a bioluminescent substrate for oxidative protein folding. Protein Sci 27:1509-1517
Fass, Deborah; Thorpe, Colin (2018) Chemistry and Enzymology of Disulfide Cross-Linking in Proteins. Chem Rev 118:1169-1198
Foster, Celia K; Thorpe, Colin (2017) Challenges in the evaluation of thiol-reactive inhibitors of human protein disulfide Isomerase. Free Radic Biol Med 108:741-749
Zhang, Han; Trout, William S; Liu, Shuang et al. (2016) Rapid Bioorthogonal Chemistry Turn-on through Enzymatic or Long Wavelength Photocatalytic Activation of Tetrazine Ligation. J Am Chem Soc 138:5978-83
Hudson, Devin A; Thorpe, Colin (2015) Mia40 is a facile oxidant of unfolded reduced proteins but shows minimal isomerase activity. Arch Biochem Biophys 579:1-7
Sapra, Aparna; Ramadan, Danny; Thorpe, Colin (2015) Multivalency in the inhibition of oxidative protein folding by arsenic(III) species. Biochemistry 54:612-21
Hudson, Devin A; Gannon, Shawn A; Thorpe, Colin (2015) Oxidative protein folding: from thiol-disulfide exchange reactions to the redox poise of the endoplasmic reticulum. Free Radic Biol Med 80:171-82
Israel, Benjamin A; Jiang, Lingxi; Gannon, Shawn A et al. (2014) Disulfide bond generation in mammalian blood serum: detection and purification of quiescin-sulfhydryl oxidase. Free Radic Biol Med 69:129-35
Schaefer-Ramadan, Stephanie; Thorpe, Colin; Rozovsky, Sharon (2014) Site-specific insertion of selenium into the redox-active disulfide of the flavoprotein augmenter of liver regeneration. Arch Biochem Biophys 548:60-5

Showing the most recent 10 out of 90 publications